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Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Volume 18 - Number 9 September 2014 The PDF version of the Atlas of Genetics and Cytogenetics in Oncology and Haematology is a reissue of the original articles published in collaboration with the Institute for Scientific and Technical Information (INstitut de l’Information Scientifique et Technique - INIST) of the French National Center for Scientific Research (CNRS) on its electronic publishing platform I-Revues. Online and PDF versions of the Atlas of Genetics and Cytogenetics in Oncology and Haematology are hosted by INIST-CNRS. Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Scope The Atlas of Genetics and Cytogenetics in Oncology and Haematology is a peer reviewed on-line journal in open access, devoted to genes, cytogenetics, and clinical entities in cancer, and cancer-prone diseases. It presents structured review articles (“cards”) on genes, leukaemias, solid tumours, cancer-prone diseases, and also more traditional review articles (“deep insights”) on the above subjects and on surrounding topics. It also present case reports in hematology and educational items in the various related topics for students in Medicine and in Sciences. Editorial correspondance Jean-Loup Huret Genetics, Department of Medical Information, University Hospital F-86021 Poitiers, France tel +33 5 49 44 45 46 or +33 5 49 45 47 67 [email protected] or [email protected] Staff Mohammad Ahmad, Mélanie Arsaban, Marie-Christine Jacquemot-Perbal, Vanessa Le Berre, Anne Malo, Carol Moreau, Catherine Morel-Pair, Laurent Rassinoux, Alain Zasadzinski. Philippe Dessen is the Database Director, and Alain Bernheim the Chairman of the on-line version (Gustave Roussy Institute – Villejuif – France). The Atlas of Genetics and Cytogenetics in Oncology and Haematology (ISSN 1768-3262) is published 12 times a year by ARMGHM, a non profit organisation, and by the INstitute for Scientific and Technical Information of the French National Center for Scientific Research (INIST-CNRS) since 2008. The Atlas is hosted by INIST-CNRS (http://www.inist.fr) http://AtlasGeneticsOncology.org © ATLAS - ISSN 1768-3262 The PDF version of the Atlas of Genetics and Cytogenetics in Oncology and Haematology is a reissue of the original articles published in collaboration with the Institute for Scientific and Technical Information (INstitut de l’Information Scientifique et Technique - INIST) of the French National Center for Scientific Research (CNRS) on its electronic publishing platform I-Revues. Online and PDF versions of the Atlas of Genetics and Cytogenetics in Oncology and Haematology are hosted by INIST-CNRS. Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Editor Jean-Loup Huret (Poitiers, France) Editorial Board Sreeparna Banerjee Alessandro Beghini Anne von Bergh Judith Bovée Vasantha Brito-Babapulle Charles Buys Anne Marie Capodano Fei Chen Antonio Cuneo Paola Dal Cin Brigitte Debuire François Desangles Enric Domingo-Villanueva Ayse Erson Richard Gatti Ad Geurts van Kessel Oskar Haas Anne Hagemeijer Nyla Heerema Jim Heighway Sakari Knuutila Lidia Larizza Lisa Lee-Jones Edmond Ma Roderick McLeod Cristina Mecucci Fredrik Mertens Konstantin Miller Felix Mitelman Hossain Mossafa Stefan Nagel Florence Pedeutour Elizabeth Petty Susana Raimondi Mariano Rocchi Alain Sarasin Albert Schinzel Clelia Storlazzi Sabine Strehl Nancy Uhrhammer Dan Van Dyke Roberta Vanni Franck Viguié José Luis Vizmanos Thomas Wan Adriana Zamecnikova (Ankara, Turkey) (Milan, Italy) (Rotterdam, The Netherlands) (Leiden, The Netherlands) (London, UK) (Groningen, The Netherlands) (Marseille, France) (Morgantown, West Virginia) (Ferrara, Italy) (Boston, Massachussetts) (Villejuif, France) (Paris, France) (London, UK) (Ankara, Turkey) (Los Angeles, California) (Nijmegen, The Netherlands) (Vienna, Austria) (Leuven, Belgium) (Colombus, Ohio) (Liverpool, UK) (Helsinki, Finland) (Milano, Italy) (Newcastle, UK) (Hong Kong, China) (Braunschweig, Germany) (Perugia, Italy) (Lund, Sweden) (Hannover, Germany) (Lund, Sweden) (Cergy Pontoise, France) (Braunschweig, Germany) (Nice, France) (Ann Harbor, Michigan) (Memphis, Tennesse) (Bari, Italy) (Villejuif, France) (Schwerzenbach, Switzerland) (Bari, Italy) (Vienna, Austria) (Clermont Ferrand, France) (Rochester, Minnesota) (Montserrato, Italy) (Paris, France) (Pamplona, Spain) (Hong Kong, China) (Kuwait) Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Solid Tumours Section Genes Section Genes / Leukaemia Sections Solid Tumours Section Leukaemia Section Deep Insights Section Solid Tumours Section Genes / Deep Insights Sections Leukaemia Section Genes / Solid Tumours Section Deep Insights Section Leukaemia / Solid Tumours Sections Solid Tumours Section Solid Tumours Section Cancer-Prone Diseases / Deep Insights Sections Cancer-Prone Diseases Section Genes / Leukaemia Sections Deep Insights Section Leukaemia Section Genes / Deep Insights Sections Deep Insights Section Solid Tumours Section Solid Tumours Section Leukaemia Section Deep Insights / Education Sections Genes / Leukaemia Sections Solid Tumours Section Education Section Deep Insights Section Leukaemia Section Deep Insights / Education Sections Genes / Solid Tumours Sections Deep Insights Section Genes / Leukaemia Section Genes Section Cancer-Prone Diseases Section Education Section Genes Section Genes / Leukaemia Sections Genes / Cancer-Prone Diseases Sections Education Section Solid Tumours Section Leukaemia Section Leukaemia Section Genes / Leukaemia Sections Leukaemia Section Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Volume 18, Number 9, September 2014 Table of contents Gene Section AFAP1L2 (actin filament associated protein 1-like 2) Xiaohui Bai, Serisha Moodley, Hae-Ra Cho, Mingyao Liu 628 ENOX2 (ecto-NOX disulfide-thiol exchanger 2) Xiaoyu Tang, Dorothy M Morré, D James Morré 632 FABP7 (fatty acid binding protein 7, brain) Roseline Godbout, Ho-Yin Poon, Rong-Zong Liu 638 GSTA1 (glutathione S-transferase alpha 1) Ana Savic-Radojevic, Tanja Radic 645 MOAP1 (Modulator Of Apoptosis 1) Gamze Ayaz, Mesut Muyan 650 PHLDA1 (pleckstrin homology-like domain, family A, member 1) Maria Aparecida Nagai 652 ADAMTS15 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif, 15) Santiago Cal, Alvaro J Obaya 655 ADRB2 (adrenoceptor beta 2, surface) Denise Tostes Oliveira, Diego Mauricio Bravo-Calderón 659 IRF4 (interferon regulatory factor 4) Vipul Shukla, Runqing Lu 663 PLCG1 (Phospholipase C, Gamma 1) Rebeca Manso 668 SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5) Cesare Indiveri, Lorena Pochini, Michele Galluccio, Mariafrancesca Scalise 673 USB1 (U6 snRNA biogenesis 1) Elisa Adele Colombo 678 Leukaemia Section t(9;15)(p13;q24) PAX5/PML Jean-Loup Huret 682 t(1;9)(p13;p12) PAX5/HIPK1 Jean-Loup Huret 685 Solid Tumour Section Lung: t(6;12)(q22;q14.1) LRIG3/ROS1 in lung adenocarcinoma Kana Sakamoto, Yuki Togashi, Kengo Takeuchi Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 688 Atlas of Genetics and Cytogenetics in Oncology and Haematology OPEN ACCESS JOURNAL INIST-CNRS Deep Insight Section The tumour suppressor function of the scaffolding protein spinophilin Denis Sarrouilhe, Véronique Ladeveze 691 Case Report Section Translocation t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia patient Adriana Zamecnikova, Soad Al Bahar, Ramesh Pandita Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 701 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Gene Section Review AFAP1L2 (actin filament associated protein 1like 2) Xiaohui Bai, Serisha Moodley, Hae-Ra Cho, Mingyao Liu Latner Thoracic Surgery Research Laboratoires, University Health Network, Toronto General Research Institute, University of Toronto, Toronto, Ontario, Canada (XB, SM, HRC, ML) Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/AFAP1L2ID52197ch10q25.html DOI: 10.4267/2042/54026 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology Abstract Identity Review on AFAP1L2, with data on DNA/RNA, on the protein encoded and where the gene is implicated. Other names: KIAA1914, XB130 HGNC (Hugo): AFAP1L2 Location: 10q25.3 Figure 1. XB130 chromosomal location and neighbour genes. A. xb130 gene is located on chromosome 10, at 10q25.3 by fluorescence in situ hybridization (FISH). B. Diagram of xb130 neighbour genes between 115939029 and 116450393. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 628 AFAP1L2 (actin filament associated protein 1-like 2) Bai X, et al. Figure 2. XB130 functional domains and motifs. Human XB130 has 818 amino acids. It contains the following motif/domains: proline-rich region: residues 98-107; tyrosine phosphorylation motif: residues 54-57, 124-127; 148-151; 457-460; PH domain: residues 175-272; 353-445; coiled-coil region: residues 652-749. approximately 130 kDa by western blotting (Xu et al., 2007). As an adaptor protein, XB130 has no enzymatic domains or activity. Sequence structure analysis has revealed 23 putative tyrosine phosphorylation sites and 27 putative phosphorylation sites for serine/threonine kinases (Xu et al., 2007). The N-terminal of XB130 contains a proline rich, SH3 domain binding motif, three tyrosine containing SH2 domain binding sites (Xu et al., 2007), of which a YXXM motif is for PI3 kinase subunit p85 binding (Lodyga et al., 2009). In the middle region, there are two pleckstrin homology domains and another tyrosine binding motif (Xu et al., 2007). The C-terminal contains a coiled-coil region, which may be important for molecular trafficking or dimerization (Xu et al., 2007). DNA/RNA Note Human XB130 was discovered in Dr. Mingyao Liu's laboratory (University of Toronto) in the process of cloning human actin filament associated protein (afap) gene. Using chicken AFAP protein sequence to search human cDNA library in GenBank, XB130 was found as an EST clone (GenBank accession number 1154093) with 34% sequence similarity to chicken AFAP protein. The clone contains partial coding sequence and 3' UTR. The upstream sequence was obtained using 5' rapid amplification of cDNA ends (RACE) from human lung alveolar epithelial cell mRNA. Western blot shows the protein molecular weight is 130 kD (Xu et al., 2007). In 2003, the XB130 knockout mice were established through the collaboration of Drs. Mingyao Liu and Tak W. Mak at the University of Toronto. Expression In normal human tissue, the 4 kb mRNA transcript of XB130 is expressed highly in spleen and thyroid with lower expression in kidney, brain, lung and pancreas (Xu et al., 2007). Newer RNA sequencing by Illumina body map using RNA obtained from 16 normal human tissues shows high expression of XB130 in thyroid with lower expression in lymph nodes, brain, colon, adipocytes, kidney, lung, adrenal glands, breast, ovary, prostate and testis followed by whole blood, heart, skeletal muscles and liver (www.genecards.org). XB130 protein is detected in normal tissues of thyroid, parathyroid, brain, kidney, skin and GItracks (www.proteinatlas.org). XB130 protein expresses in human thyroid, colorectal, gastric and hepatocellular carcinomas (Shi et al., 2012; Shiozaki et al., 2013; Shiozaki et al., 2011; Zuo et al., 2012). Expression of XB130 has also been observed in a variety of cancer cell lines, including thyroid, lung, esophageal, pancreatic and colon cancers (Shi et al., 2012; Shiozaki et al., 2013; Zuo et al., 2012). Description Human xb130 genes contains 19 exons, which are covering the whole coding sequence. Transcription The transcript size of xb130 is 3751 bp. There may be 7 splicing variants based on Ensembl data (www.ensembl.org). XB130 mRNA is highly expressed in the thyroid, parathyroid and spleen; moderately expressed in brain, pancreas, lung and kidney. Protein Description XB130 is a novel adaptor protein, member of the actin filament associated protein (AFAP) family (Snyder et al., 2011). Accordingly, it is also known as AFAP1L2. The full length protein consists of 818 amino acids with a molecular weight of Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 629 AFAP1L2 (actin filament associated protein 1-like 2) Bai X, et al. lung, large intestine, ovary, skin, prostate, endometrium. Among these samples, 70% of identified cases are XB130 substitution missense mutation. Localisation XB130 is distributed mainly in the cytoplasm and perinuclear region of lung epithelial BEAS-2B cells and several other cell types (Xu et al., 2007). Unlike AFAP, XB130 does not associate or colocalize with actin filament stress fibler (Lodyga et al., 2010). Stimulation of cells with EGF, PMA, or overexpression of constitutive Rac results in a translocation of XB130 to the cell periphery and leading edge of migrating cells (Lodyga et al., 2010). Implicated in Various cancers Note XB130 plays important roles in tumor progression by promoting cell proliferation, survival, motility and invasion in various cancer cells. Recently, XB130 has been identified in thyroid carcinoma (Shiozaki et al., 2011), esophageal squamous cell carcinoma (Shiozaki et al., 2013), and gastric cancer (Shi et al., 2012). Function XB130 is an adaptor protein that acts as a key mediator to drive signal transduction pathways. XB130 has been shown to bind to tyrosine kinase cSrc to enhance kinase activity and subsequently regulates Src-mediated AP-1/SRE transcription activation (Xu et al., 2007). XB130 is also highly involved in the PI3K/Akt pathway and effects cell proliferation, cell cycle progression and cell survival through binding to p85 alpha subunit of PI3K (Lodyga et al., 2009). XB130 may also play a role in the innate immune response, where knockdown of XB130 was shown to decrease IL-6 and IL-8 cytokine levels in human lung epithelial cells (Xu et al., 2007). XB130 is also involved in cell migration via association with RacGTPase and plays a significant role in lateral cell migration and cell invasion in both normal and cancer cell lines (Lodyga et al., 2010). Yamanaka et al. reported phosphorylated XB130 affects cAMP-dependent DNA synthesis in rat thyroid cells (Yamanaka et al., 2012). XB130 is aberrantly expressed in human cancers and has been shown to control tumour growth in vivo (Shiozaki et al., 2011). XB130 regulates thyroid cancer cell proliferation by controlling microRNA miR-33a, 149a and 193a expression to alter oncogenes Myc, FosL1, and SCL7A5 protein levels (Takeshita et al., 2013). Colorectal cancer Note Tyrosine phosphorylated XB130 in colorectal cancer. Prognosis Using mass spectrometry, Emaduddin et al. reported several proteins as tyrosine phosphorylated form are maintained at high level in colorectal cancer cells isolated from patients. XB130 is identified as one of these proteins. Therefore, tyrosine phosphorylated XB130 has a potential to be a biomarker of colorectal cancer (Emaduddin et al., 2008). Gastric cancer (GC) Note XB130 expression level associates with the prognosis of gastric cancer. Prognosis Based on the anlysis GC tissue samples from 411 patients with various stages, lower expression of XB130 mRNA as well as protein is significantly correlated with reduced patient survival time and shorter disease-free period (Shi et al., 2012). Homology Thyroid cancer XB130 shares similar sequence and domain structure cellular as AFAP and AFAP1L1 (Snyder et al., 2011). Note XB130 as a tumor promoting gene, enhancing thyroid cancer cell growth. Oncogenesis Knockdown XB130 using siRNA in thyroid cancer cell (WRO) is accompanied with an inhibition of G1-S phase cell cycle progression and enhanced apoptosis. The volume of tumors generated in nude mice after injecting these cells are smaller than those formed from cells with a normal XB130 expression (Shiozaki et al., 2011). Mutations Somatic Somatic mutations of XB130 are reported in a variety of cancer tissues. Based on the data of Sanger Institute database (www.sanger.ac.uk), XB130 mutation sites have been identified in multiple tumor tissues, including Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 630 AFAP1L2 (actin filament associated protein 1-like 2) Bai X, et al. Esophageal squamous cell carcinoma (ESCC) is a Rac-controlled component of lamellipodia that regulates cell motility and invasion. J Cell Sci. 2010 Dec 1;123(Pt 23):4156-69 Note XB130 protein is identified in ESCC primary cell lines and tumor samples. Oncogenesis XB130 protein is highly expression in ESCC primary cells. XB130 protein is examined by immunohistochemistry staining from ESCC tissues collected from 52 patients. Positive XB130 staining is observed in 71% of ESCC samples, which indicates the association of XB130 protein and ESCC (Shiozaki et al., 2013). Shiozaki A, Liu M. Roles of XB130, a novel adaptor protein, in cancer. J Clin Bioinforma. 2011 Mar 17;1(1):10 Shiozaki A, Lodyga M, Bai XH, Nadesalingam J, Oyaizu T, Winer D, Asa SL, Keshavjee S, Liu M. XB130, a novel adaptor protein, promotes thyroid tumor growth. Am J Pathol. 2011 Jan;178(1):391-401 Snyder BN, Cho Y, Qian Y, Coad JE, Flynn DC, Cunnick JM. AFAP1L1 is a novel adaptor protein of the AFAP family that interacts with cortactin and localizes to invadosomes. Eur J Cell Biol. 2011 May;90(5):376-89 Shi M, Huang W, Lin L, Zheng D, Zuo Q, Wang L, Wang N, Wu Y, Liao Y, Liao W. Silencing of XB130 is associated with both the prognosis and chemosensitivity of gastric cancer. PLoS One. 2012;7(8):e41660 Soft tissue tumor Note Decreased XB130 expression leads to a local aggressiveness of soft tissue tumor. Oncogenesis Analysis of gene expression profile of 102 tumor samples with varying stages of soft tissue tumor shows a decreased XB130 expression in malignant mesenchymal tumors (Cunha et al., 2010). Shiozaki A, Shen-Tu G, Bai X, Iitaka D, De Falco V, Santoro M, Keshavjee S, Liu M. XB130 mediates cancer cell proliferation and survival through multiple signaling events downstream of Akt. PLoS One. 2012;7(8):e43646 References Zuo Q, Huang H, Shi M, Zhang F, Sun J, Bin J, Liao Y, Liao W. Multivariate analysis of several molecular markers and clinicopathological features in postoperative prognosis of hepatocellular carcinoma. Anat Rec (Hoboken). 2012 Mar;295(3):423-31 Yamanaka D, Akama T, Fukushima T, Nedachi T, Kawasaki C, Chida K, Minami S, Suzuki K, Hakuno F, Takahashi S. Phosphatidylinositol 3-kinase-binding protein, PI3KAP/XB130, is required for cAMP-induced amplification of IGF mitogenic activity in FRTL-5 thyroid cells. Mol Endocrinol. 2012 Jun;26(6):1043-55 Xu J, Bai XH, Lodyga M, Han B, Xiao H, Keshavjee S, Hu J, Zhang H, Yang BB, Liu M. XB130, a novel adaptor protein for signal transduction. J Biol Chem. 2007 Jun 1;282(22):16401-12 Shiozaki A, Kosuga T, Ichikawa D, Komatsu S, Fujiwara H, Okamoto K, Iitaka D, Nakashima S, Shimizu H, Ishimoto T, Kitagawa M, Nakou Y, Kishimoto M, Liu M, Otsuji E. XB130 as an independent prognostic factor in human esophageal squamous cell carcinoma. Ann Surg Oncol. 2013 Sep;20(9):3140-50 Emaduddin M, Edelmann MJ, Kessler BM, Feller SM. Odin (ANKS1A) is a Src family kinase target in colorectal cancer cells. Cell Commun Signal. 2008 Oct 9;6:7 Lodyga M, De Falco V, Bai XH, Kapus A, Melillo RM, Santoro M, Liu M. XB130, a tissue-specific adaptor protein that couples the RET/PTC oncogenic kinase to PI 3-kinase pathway. Oncogene. 2009 Feb 19;28(7):937-49 Takeshita H, Shiozaki A, Bai XH, Iitaka D, Kim H, Yang BB, Keshavjee S, Liu M. XB130, a new adaptor protein, regulates expression of tumor suppressive microRNAs in cancer cells. PLoS One. 2013;8(3):e59057 Cunha IW, Carvalho KC, Martins WK, Marques SM, Muto NH, Falzoni R, Rocha RM, Aguiar S, Simoes AC, Fahham L, Neves EJ, Soares FA, Reis LF. Identification of genes associated with local aggressiveness and metastatic behavior in soft tissue tumors. Transl Oncol. 2010 Feb;3(1):23-32 This article should be referenced as such: Bai X, Moodley S, Cho HR, Liu M. AFAP1L2 (actin filament associated protein 1-like 2). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):628-631. Lodyga M, Bai XH, Kapus A, Liu M. Adaptor protein XB130 Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 631 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Gene Section Review ENOX2 (ecto-NOX disulfide-thiol exchanger 2) Xiaoyu Tang, Dorothy M Morré, D James Morré MorNuCo, Inc., 1201 Cumberland Avenue, Ste. B, Purdue Research Park,.West Lafayette, IN 47906 USA (XT, DMM, DJM) Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/ENOX2ID40134chXq26.html DOI: 10.4267/2042/54027 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology according to NCBI Refseq Gene Database (gene ID: 10495, RefSeq ID: NG_012562.1), and is comprised of 279939 bp. ENOX2 is composed of 13 protein-coding exons between 71 bp and 2066 bp in length and 14 introns which vary greatly in length (1781 bp to 117994 bp). It has a 501 bp 5' untranslated region and a long 3' UTR (approximately 1935 bp). Abstract Review on ENOX2, with data on DNA/RNA, on the protein encoded and where the gene is implicated. Identity Other names: APK1, COVA1, tNOX HGNC (Hugo): ENOX2 Location: Xq26.1 Note Also termed APK1 antigen, or cytosolic ovarian carcinoma antigen 1, or tumor-associated hydroquinone oxidase (tNOX). ECTO-NOX2 = Ecto-Nicotinamide Dinucleotide Oxidase Disulfide Thiol Exchange 2. Transcription According to NCBI the human ENOX2 gene encodes a 4036 bp mRNA transcript, the coding sequence (CDS) located from base pairs 356 to 2101 (NM_001281736.1). The CDS from the Ensembl genome browser database (ENST00000370927, transcript length 3788 bp) and NCBI (NM_001281736.1) are identical. Transcripts NM_001281736.1 and ENST 00000370927 are also included in the human CCDS set (CCDS14626) and encode a 610 aa long protein. DNA/RNA Description Pseudogene The human ENOX2 gene is located on the reverse strand of chromosome X (bases 4918 to 284856); None known. Figure 1. ENOX2 mRNA. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 632 ENOX2 (ecto-NOX disulfide-thiol exchanger 2) Tang X, et al. Figure 2. Deduced amino acid sequence and functional motifs of the bacterially expressed 46 kDa enzymatically active Cterminus of ENOX2. for protein disulfide-thiol interchange (Morré and Morré, 2013). The signature ENOX2 motif is that of the potential drug/antibody binding site E394EMTE. Antisera directed to this portion of the protein act as competitive inhibitors to drug binding. The sequence provides a putative quinone or sulfonylurea-binding site with four of the five amino acids in at least one other putative quinone site in the same relative positions. The correctness of the various assignments has, for the most part, been confirmed by site-directed mutagenesis (Chueh et al., 2002). While amino acid replacements that block oxidation of reduced pyridine nucleotide by ENOX2 also eliminated protein disulfide-thiol interchange and vice versa (Chueh et al., 2002), the two activities appear to occur independently. One can be measured in the absence of the other. The ENOX2 proteins have properties of prions and are protease resistant (Kelker et al., 2001) and Nterminal sequencing. Concentrated solutions aggregate and form amyloid-like filaments. Protein Description ENOX2 transcription variants all appear to be variations that include an exon 4 minus splicing event that allows for down-stream initiation and expression of the ENOX2 protein at the cell surface of malignant cells (Tang et al., 2007a; Tang et al., 2007b). Without the exon 4 deletion, mRNA derived from the gene does not appear to be translated into protein. Thus, the exon 4 deletion is the basis for the cancer specificity of the ENOX2 transcription variants. An hnRNP splicing factor directs formation of the Exon 4 minus variants of ENOX2 (Tang et al., 2011). The fully processed 34 kDa generic ENOX2 protein found on the cell surface of HeLa cells and in sera of about 23% of early cancer patients retains full-functional activity. The deduced amino acid sequence of a bacterially expressed 46 kDa functional C-terminus of ENOX2 exhibits the same characteristics of alternation of the two activities and drug response as the cell surface and generic serums forms. Identified functional motifs include a quinone binding site, an adenine nucleotide binding site, a CXXXXC cysteine motif as a potential disulfide-thiol interchange site and two copper binding sites, one of which is conserved with superoxide dismutase. ENOX2 proteins lack flavin and only one of the two C-X-X-X-X-C motifs characteristic of flavoproteins are present in ENOX2. Yet the protein effectively carries out protein disulfide interchange. The motif C569-X-X-X-X-X-C575, alone or together with a downstream histidine (H582) provides an additional potential active site Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Expression Widely expressed in malignant cells but only as exon 4 minus splicing variants (Tang et al., 2007b). Localisation External cell surface (Morré, 1995). Function ENOX2 is a member of a family of cell surface metalocatalysts with binuclear copper centers that oscillate. 633 ENOX2 (ecto-NOX disulfide-thiol exchanger 2) Tang X, et al. Figure 3. Diagrammatic representation of the functional unit of the ENOX2 proteins which is a dimer, each monomer of which contains two copper centers. During the oxidative portion of the ENOX cycle on the right, the net result is the transfer of 4 electrons plus 4 protons to molecular oxygen to from 2H2O. The left portion of the diagram illustrates the protein disulfide-thiol interchange activity portion of the cycle where the result is an interchange of protons and electrons that results in the breakage and formation of disulfide bonds important to cell enlargement. They catalyze both NAD(P)H and hydroquinone oxidation in one configuration and carry out protein disulfide-thiol interchange in a second configuration (Figure 3). The two activities alternate creating a regular 22 min period to impart a time-keeping function (Morré and Morré, 2003). The oscillations are highly synchronous and phased by low frequency electromagnetic fields. Functionally ENOX2 proteins of cancer cells act as terminal oxidases of plasma membrane electron transport (PMET) whereby electrons coming from cytosolic NAD(P)H are transferred to membranelocated coenzyme Q with eventual transfer of electrons and proteins to oxygen to form water (Figure 4). The released energy is presumably utilized to drive cell enlargement. The protein disulfide-thiol interchange part of the cycle carries out a function essential to the cell enlargement mechanism (Morré et al., 2006). The phenotype of unregulated accelerated growth is recapitulated in a transgenic mouse strain over expressing human ENOX2 (Yagiz et al., 2006). The ENOX2 gene is present in the human genome as a single copy, with no obvious homologs and a single constitutive ENOX1 (CNOX) ortholog with 64% identity and 80% similarity (Jiang et al., 2008). Mutations Somatic No reports of mutations leading to inactivation of or inability to express ENOX2. Implicated in Various cancers Note The ENOX2 protein is universally associated with malignancies. It is not the result of an oncogenic mutation but appears to be similar if not identical to a form of ENOX protein with characteristics of an oncofetal protein important to maintenance of unregulated growth in very early development that may be re-expressed in malignancy (Cho and Morré, 2009). Re-expression as an oncofetal protein helps explain the role of ENOX2 of cancer cells in acquiring the well-known characteristic of uncontrolled growth. Consistent with this interpretation are observations that the malignant phenotypes of invasiveness and growth on soft agar of cancer cells in culture are lost when cells are transfected with ENOX2 antisense (Chueh et al., 2004; Tang et al., 2007a). Homology RNA recognition motif (RRM) in the cell surface Ecto-NOX disulfide-thiol exchanger (ECTO-NOX or ENOX) proteins. This subgroup corresponds to the conserved RNA recognition motif (RRM) in ECTO-NOX proteins (also termed ENOX), comprising a family of plant and animal NAD(P)H oxidases exhibiting both oxidative and protein disulfide-like activities. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 634 ENOX2 (ecto-NOX disulfide-thiol exchanger 2) Tang X, et al. Figure 4. Schematic representation of the utility of the ENOX2 proteins as cancer-specific cell surface proteins for diagnosis and therapeutic intervention in cancer. Modified from Morré and Morré (2013). ENOX2 is the first reported cell surface change absent from non-cancer cells and associated with most, if not all, forms of human cancer (Morré and Morré, 2013). As such, ENOX2 emerges as a potential universal molecular cancer marker and, being an ecto-protein at the cell surface and shed into the circulation, a reliable cancer diagnostic marker both for cancer presence and tissue of cancer origin (Figure 4). ENOX2 proteins are expressed differently by different tissues of cancer origin within the body with each type of cancer being characterized by one, two, three or more tissue specific transcript variants of characteristic molecular weights and isoelectric points (Morré and Morré, 2012). ENOX2 proteins are absent or reduced to below the limits of detection from sera of healthy individuals or patients with diseases other than cancer. Circulating ENOX2 has been detected in sera of patients representing all major forms of human cancer including leukemias and lymphomas. All ENOX2 transcript variants appear to share the common antigenic determinant recognized both by an ENOX2-specific monoclonal antibody (Cho et al., 2002) and a corresponding scFv single chain variable region recombinant antibody expressed in bacteria and derived from the monoclonal antibodyproducing hybridoma cells with analysis by 2-D-gel electrophoresis and western blot (Hostetler et al., 2009). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Breast cancer Note Sera of breast cancer patients contains an ENOX2 transcript variant of 64 to 69 kDa, isoelectric point 4.2 to 4.9. Lung cancer Note Sera of patients with non-small cell lung cancer contain a 53 to 56 kDa ENOX2 transcript variant, isoelectric point pH 4.7 to 5.3 while sera of small cell lung cancer contain a transcript variant of 52 kDa, isoelectric point pH 4.1 to 4.6. Prostate cancer Note Sera of patients with prostate cancer contain a 71 to 88 kDa ENOX2 transcript variant, isoelectric point pH 5.1 to 6.5. Cervical cancer Note Sera of cervical cancer patients contain a 90 to 100 kDa transcript variant, isoelectric point pH 4.2 to 5.4. Malignant melanoma Note Sera from malignant melanoma patients contain an ENOX2 transcript variant of 37 to 41 kDa, 635 ENOX2 (ecto-NOX disulfide-thiol exchanger 2) Tang X, et al. isoelectric point pH 4.6 to 5.3. by anticancer drugs such as doxorubicin, the anticancer sulfonylureas, the vanilloid capsaicin, the catechin EGCg and the cancer isoflavene phenoxodiol, all of which appear to function as quinone site inhibitors directed toward the EEMTE drug binding motif of ENOX2 (Morré and Morré, 2013; Hanau et al., 2014). The possibility of ENOX2 as a drug target is enhanced by the external location of the ENOX2 protein in a position to be readily available to drugs or antibodies conjugated to impermeant supports. As the growth involvement of ENOX2 proteins is in cell enlargement, ENOX2 inhibitors also block cell proliferation. The blocked cells, unable to enlarge, also fail to divide and eventually undergo apoptosis (Figure 4). Leukemias, lymphomas and myelomas Note Sera of patients with leukemia, lymphoma or myeloma, cancers having blood as the common tissue of origin, all contain ENOX2 transcript variants of 38 to 48 kDa and low isoelectric point pH 3.6 to 4.5. Ovarian cancer Note Sera from ovarian carcinoma patients contain two ENOX2 transcript variants of 72 to 90 kDa and 37 to 47 kDa, both having similar isoelectric points in the range of pH 3.7 to 5.0. References Bladder cancer Morré DJ. NADH oxidase activity of HeLa plasma membranes inhibited by the antitumor sulfonylurea N-(4methylphenylsulfonyl)-N'-(4-chlorophenyl) urea (LY181984) at an external site. Biochim Biophys Acta. 1995 Dec 13;1240(2):201-8 Note Sera of patients with carcinoma of the bladder contain two ENOX2 transcript variants of 63 to 66 kDa and 42 to 48 kDa with isoelectric points of 4.2 to 5.8 and 4.1 to 4.8, respectively. Kelker M, Kim C, Chueh PJ, Guimont R, Morré DM, Morré DJ. Cancer isoform of a tumor-associated cell surface NADH oxidase (tNOX) has properties of a prion. Biochemistry. 2001 Jun 26;40(25):7351-4 Uterine cancer Note Sera of patients with uterine carcinoma contain two ENOX2 transcript variants of 64 to 69 kDa and 36 to 48 kDa with isoelectric points of pH 4.2 to 4.9 and pH 4.5 to 5.6. Cho N, Chueh PJ, Kim C, Caldwell S, Morré DM, Morré DJ. Monoclonal antibody to a cancer-specific and drugresponsive hydroquinone (NADH) oxidase from the sera of cancer patients. Cancer Immunol Immunother. 2002 May;51(3):121-9 Chueh PJ, Kim C, Cho N, Morré DM, Morré DJ. Molecular cloning and characterization of a tumor-associated, growth-related, and time-keeping hydroquinone (NADH) oxidase (tNOX) of the HeLa cell surface. Biochemistry. 2002 Mar 19;41(11):3732-41 Colorectal cancer Note Sera of patients with colorectal cancer contain at least two of three possible ENOX2 transcript variants of 80 to 96 kDa, isoelectric point pH 4.5 to 5.3, 50 to 60 kDa, isoelectric point pH 4.2 to 5.1 and 33 to 46 kDa, isoelectric point pH 3.8 to 5.2. Morré DJ, Morré DM. Cell surface NADH oxidases (ECTONOX proteins) with roles in cancer, cellular time-keeping, growth, aging and neurodegenerative diseases. Free Radic Res. 2003 Aug;37(8):795-808 Other cancers Chueh PJ, Wu LY, Morré DM, Morré DJ. tNOX is both necessary and sufficient as a cellular target for the anticancer actions of capsaicin and the green tea catechin (-)-epigallocatechin-3-gallate. Biofactors. 2004;20(4):23549 Note Unique patterns of ENOX2 transcript variant expression (number, molecular with and isoelectric point) have been found as well associated with brain, endometrial, esophageal, gastric, hepatocellular renal cell, squamous cell, testicular germ cell and thyroid cancer as well as mesothelioma and sarcomas. Morré DJ, Kim C, Hicks-Berger C. ATP-dependent and drug-inhibited vesicle enlargement reconstituted using synthetic lipids and recombinant proteins. Biofactors. 2006;28(2):105-17 Yagiz K, Morré DJ, Morré DM. Transgenic mouse line overexpressing the cancer-specific tNOX protein has an enhanced growth and acquired drug-response phenotype. J Nutr Biochem. 2006 Nov;17(11):750-9 Endometriosis Note Invasive endometriosis is the only non-malignant disorder thus far characterized by the presence of unique ENOX2 transcript variants. Tang X, Morré DJ, Morré DM. Antisense experiments demonstrate an exon 4 minus splice variant mRNA as the basis for expression of tNOX, a cancer-specific cell surface protein. Oncol Res. 2007a;16(12):557-67 As a cancer therapeutic drug target Tang X, Tian Z, Chueh PJ, Chen S, Morré DM, Morré DJ. Alternative splicing as the basis for specific localization of tNOX, a unique hydroquinone (NADH) oxidase, to the cancer cell surface. Biochemistry. 2007b Oct 30;46(43):12337-46 Note ENOX2 is responsive to differentiating agents such as calcitriol and anticancer retinoids and inhibited Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 636 ENOX2 (ecto-NOX disulfide-thiol exchanger 2) Tang X, et al. Jiang Z, Gorenstein NM, Morré DM, Morré DJ. Molecular cloning and characterization of a candidate human growthrelated and time-keeping constitutive cell surface hydroquinone (NADH) oxidase. Biochemistry. 2008 Dec 30;47(52):14028-38 Morre DJ, Morre DM.. Early detection: an opportunity for cancer prevention through early intervention. In: Georgakilas AG (ed), Cancer Prevention. http://www.intechopen.com/download/pdf/35600. In Tech, Rijeka, 2012:389-402 pp. Cho N, Morré DJ. Early developmental expression of a normally tumor-associated and drug-inhibited cell surfacelocated NADH oxidase (ENOX2) in non-cancer cells. Cancer Immunol Immunother. 2009 Apr;58(4):547-52 Morre DJ, Morre DM.. ECTO-NOX Proteins. ISBM 978-14614-3957-8. Springer, New York, 2013, 507 pp. Hanau C, Morre DJ, Morre DM.. Cancer prevention trial of a synergistic mixture of green tea concentrate plus Capsicum (CAPSOL-T) in a random population of subjects ages 40-84. http://link.springer.com/content/pdf/10.1007%252Fs12014008-9016-x. Clin Proteomics 2014: 11(2). Hostetler B, Weston N, Kim C, Morre DM, Morre DJ.. Cancer site-specific isoforms of ENOX2 (tNOX), a cancerspecific cell surface oxidase. http://link.springer.com/content/pdf/10.1007%252Fs12014008-9016-x. Clin Proteomics 2009:5:46-51. This article should be referenced as such: Tang X, Kane VD, Morre DM, Morre DJ.. hnRNP F directs formation of an exon 4 minus variant of tumor-associated NADH oxidase (ENOX2). Mol Cell Biochem. 2011 Nov;357(1-2):55-63. doi: 10.1007/s11010-011-0875-5. Epub 2011 May 28. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Tang X, Morré DM, Morré DJ. ENOX2 (ecto-NOX disulfidethiol exchanger 2). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):632-637. 637 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Gene Section Review FABP7 (fatty acid binding protein 7, brain) Roseline Godbout, Ho-Yin Poon, Rong-Zong Liu Department of Oncology, University of Alberta, Cross Cancer Institute, 11560 University Avenue, Edmonton, Alberta, T6G 1Z2 Canada (RG, HYP, RZL) Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/FABP7ID46256ch6p22.html DOI: 10.4267/2042/54028 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology Local order: PKIB → FABP7 → SMPDL3A. Abstract DNA/RNA Review on FABP7, with data on DNA/RNA, on the protein encoded and where the gene is implicated. Description The FABP7 gene is 4,5 kb long and contains 4 exons, all of which contain coding sequences. The following FABP7 SNPs have been validated: 7 in the 3' UTR, 6 in the 5' UTR, 5 missense and 4 SNPs in the coding region that don't alter the amino acid code (dbSNP). Identity Other names: B-FABP, BLBP, FABPB, MRG HGNC (Hugo): FABP7 Location: 6q22.31 FABP7 gene. The FABP7 gene is located on chromosome 6 in the region of q22-q23 on the positive strand. Neighboring genes are indicated. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 638 FABP7 (fatty acid binding protein 7, brain) Godbout R, et al. Crystal structure of FABP7 bound to DHA. The structure of FABP7 is similar to that of other FABPs and consists of two Nterminal α-helices attached to a β-barrel motif (left). Three amino acids are predicted to be important for fatty acid binding: F104 (fuchsia), arginine 126 (red) and Y128 (teal) based on the structure of FABP7 bound to DHA. DHA is shown in yellow (right). Structural data were obtained from the Protein Data Bank (PDB ID: 1FE3) (Balendiran et al., 2000) and rendered using PyMOL (Beaulieu, 2012). Based on EST data, FABP7 RNA is most highly expressed in the fetus, followed by adult brain, eye, connective tissue, bone, heart, kidney, mammary gland, skin, uterus, lung and testis. Transcription regulators: Members of the nuclear factor I (NFI) family regulate the transcription of the FABP7 gene (Bisgrove et al., 2000; Brun et al., 2009). The phosphorylation state of NFI determines its regulatory activity, with FABP7 transcription upregulated by hypophosphorylated NFI (Bisgrove et al., 2000; Brun et al., 2013). Other transcription factors implicated in the regulation of FABP7 include Notch (Anthony et al., 2005), PAX6 (Arai et al., 2005; Numayama-Tsuruta et al., 2010; Liu et al., 2012b), and POU-domain protein PBX-1 (Josephson et al., 1998). Furthermore, ligands of peroxisome proliferatoractivated receptors (PPARs) such as clofibrate and omega-3 docosahexaenoic acid (DHA) have been shown to up-regulate FABP7 expression (Nasrollahzadeh et al., 2008; Venkatachalam et al., 2012). Post-transcriptional regulation: The 3' untranslated region of FABP7 contains phylogenetically conserved cytoplasmic polyadenylation elements (CPE) which have been implicated in the trafficking and localized translation of FABP7 at perisynaptic processes of astrocytic cells (Gerstner et al., 2012). Protein Description FABP7 is a member of the intracellular lipidbinding protein family. FABP7 is a 132 amino acid polypeptide with an estimated molecular mass of 15 kDa. It has a beta-clam structure made up of ten antiparallel beta sheets capped by two alpha helices. Fatty acid ligands reside inside the beta-clam structure. Expression FABP7 is expressed in radial glial cells during brain development (Feng et al., 1994). FABP7 persists in specific regions of the mature mouse brain, including glia limitans, in radial glial cells of the hippocampal dentate gyrus and Bergman glial cells (Kurtz et al., 1994). FABP7 is also expressed in glial cells of the peripheral nervous system, and ensheathing cells of the olfactory nerve (Kurtz et al., 1994). Localisation The FABP7 protein is found in both the cytoplasm and nucleus of normal radial glial cells (Feng et al., 1994) and tumor cells (Liang et al., 2006; Slipicevic et al., 2008). FABP7 is also found in perisynaptic processes of astrocytes with localized translation of FABP7 at these sites (Gerstner et al., 2012). Pseudogene Function A predicted FABP7 pseudogene is located on chromosome 1 (NCBI nucleotide database NG_029025.1). There are two inferred human FABP7 pseudogenes listed in the Rat Genome Database (http://rgd.mcw.edu/rgdweb/report/gene/main.html? id=5132511 on chromosome 1 and http://www.rgd.mcw.edu/rgdweb/report/gene/main. html?id=6481032 on chromosome 2). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Recombinant human FABP7 exhibits the highest affinity for the polyunsaturated omega-3 fatty acids α-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, and for monounsaturated omega-9 oleic acid (Kd from 28 to 53 nM) and moderate affinity for the polyunsaturated omega-6 fatty acids, linoleic acid and arachidonic acid (AA) (Kd from 115 to 206 nM) (Balendiran et al., 2000). 639 FABP7 (fatty acid binding protein 7, brain) Godbout R, et al. FABP7 has low binding affinity for saturated long chain fatty acids. Human FABP7 enhances DHA trafficking to the nucleus (Mita et al., 2010). FABP7 is required for the establishment of the radial glial fiber system along which neurons migrate in order to reach their correct destination in the developing brain (Feng et al., 1994). FABP7 is also required for the maintenance of neuroepithelial cells in rat cortex (Arai et al., 2005). FABP7 knockout mice have a structurally normal brain; however, the mice show enhanced anxiety and increased fear memory, as well as decreased DHA in neonatal brain and increased AA in adult brain amygdala (Owada et al., 2006). increased cell migration (Liang et al., 2005). A role for FABP7 in malignant glioma cell migration was confirmed by Mita et al. (2007) who used human U87 malignant glioma cell lines stably transfected with a FABP7 expression construct to demonstrate a correlation between FABP7 expression and increased cell migration. In agreement with a role for FABP7 in migration and infiltration, FABP7 was found to be preferentially expressed at sites of infiltration and surrounding blood vessels in glioblastoma multiforme (Mita et al., 2007). Growth of malignant glioma cell lines in the presence of polyunsaturated fatty acids omega-3 DHA and omega-6 AA indicates that the ratio of AA:DHA affects migration in FABP7-positive cells, with a higher DHA:AA ratio resulting in decreased migration (Mita et al., 2010). These results suggest that glioblastoma tumour growth and infiltration may be controlled by increasing levels of DHA in tumour tissue (Elsherbiny et al., 2013). Neurospheres derived from glioblastoma multiforme express high levels of FABP7, suggesting the presence of FABP7-positive neural stem-like cells in glioblastoma (De Rosa et al., 2012). In keeping with this possibility, FABP7 is preferentially expressed in the subset of glioblastoma tumour cells that express the neural stem cell marker CD133 (Liu et al., 2009). Knockdown of FABP7 in glioblastoma-derived neurosphere cultures results in decreased cell migration and reduced proliferation (De Rosa et al., 2012). The FABP7 promoter has been shown to be hypomethylated in glioblastoma tumours compared to normal brain (Etcheverry et al., 2010). Homology Human FABP7 amino acid sequence is 86,4% identical to mouse FABP7, 90,9% identical to chicken FABP7, 82,6% identical to zebrafish FABP7a and 78% identical to zebrafish FABP7b. Human FABP7 shows variable sequence identity with the other FABP paralogues, with the lowest identity to FABP1 (27,6%) and highest identity to FABP3 (65,9%). Mutations Note With the exception of SNPs, no mutations in the FABP7 gene have been reported. Implicated in Malignant glioma (grades III and IV astrocytoma) / glioblastoma multiforme (grade IV astrocytoma) Note FABP7 was first reported to be expressed in malignant glioma cell lines and malignant glioma tumour tissue in 1998 (Godbout et al., 1998). Liang et al. (2005) used gene expression profiling to demonstrate that FABP7 RNA levels were elevated in glioblastoma tumours compared to normal brain. These authors showed that elevated levels of nuclear FABP7 protein were associated with decreased survival in patients with glioblastoma multiforme, particularly in younger patients. Subsequent analysis of 123 glioblastomas by Kaloshi et al. (2007) revealed a correlation between nuclear FABP7, EGFR amplification and more invasive tumours. De Rosa et al. (2012) also showed a correlation between elevated FABP7 levels and decreased survival in patients with glioblastoma multiforme. Transfection of a FABP7 expression construct into the SF767 malignant glioma cell line results in Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Breast cancer Note MRG (mammary-derived growth inhibitor-related gene), later shown to be identical to FABP7 (Hohoff and Spener, 1998), was reported to be expressed in normal and benign breast tissue but only rarely in breast cancer (1 of 10 infiltrative breast cancers and 2 of 12 ductal carcinomas in situ) (Shi et al., 1997). Transfection of a MRG expression construct into the MDA-MB-231 breast cancer cell line suppressed cell proliferation and tumour growth in an orthotopic mouse model (Shi et al., 1997). Subsequent work showed that MRG over-expression induced differentiation in human breast cancer cells and that treatment of breast cancer cells with DHA causes MRG-dependent growth inhibition (Wang et al., 2000). 640 FABP7 (fatty acid binding protein 7, brain) Godbout R, et al. FABP7 mRNA levels are upregulated in human glioblastoma. Comparison of FABP7 mRNA levels in normal human brain versus glioblastoma tissues. Database obtained from Oncomine website (www.oncomine.org; Bredel Brain 2). ligands available to nuclear receptors such as PPARs, understanding the roles of cytoplasmic and nuclear FABP7 will help elucidate its biological functions in breast cancer. Preferential expression of FABP7 in estrogen receptor-negative breast cancer compared to estrogen receptor-positive breast cancer has been reported by four separate groups (Tang et al., 2010; Zhang et al., 2010; Graham et al., 2011; Liu et al., 2012a). In an analysis of 176 primary breast cancers, Liu et al. (2012) found a correlation between elevated FABP7 levels and poor prognosis. These authors further showed that depletion of FABP7 in FABP7-positive/estrogennegative MDA-MB-435S, reduced cell growth and sensitized the cells to growth inhibition by DHA. In addition, FABP7 was found to mediate DHAinduced retinoid-X-receptor beta (RXRβ) activation in triple-negative BT-20 breast cancer cells as well as MDA-MB-435S cells. In a study of 899 invasive breast cancer cases, Zhang et al. (2010) showed that basal breast cancers (estrogen/progesterone receptor-negative, HER2-negative) that were FABP7-positive had significantly better outcomes than basal breast cancers that were FABP7negative. Analysis of the subcellular localization of FABP7 in 1249 unselected and 245 estrogen receptor-negative invasive breast cancers revealed both nuclear and cytoplasmic staining patterns, with nuclear FABP7 associated with a high histological grade, stage, mitotic frequency, as well as basal and triple-negative status (Alshareeda et al., 2012). Within the basal category, elevated levels of nuclear FABP7 were associated with longer disease-free survival. In light of the proposed roles for nuclear FABPs in making their fatty acid Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Renal cell carcinoma Note FABP7 RNA and protein are up-regulated in renal cell carcinoma compared to normal kidney tissue (Seliger et al., 2005; Teratani et al., 2007; Domoto et al., 2007). Analysis of a tissue microarray containing 272 renal cell carcinomas showed significantly lower levels of FABP7 in grades 3 and 4 compared to grades 1 and 2 renal cell carcinomas (Tölle et al., 2009). No correlation was found between patient survival and FABP7 staining intensity. In agreement with malignant glioma experiments, knock-down of FABP7 in human kidney carcinoma cells resulted in decreased cell migration (Tölle et al., 2011). The regulation of FABP7 in renal cell carcinoma has been addressed by analysing the FABP7 promoter (Takaoka et al., 2011). This analysis indicates that BRN2 (POU3F2) and nuclear factor I (NFI) may be regulating the expression of FABP7 in renal cell carcinoma. Melanoma Note FABP7 been reported to be both down-regulated in melanoma compared to benign nevi (de Wit et al., 2005) and widely expressed in melanoma (Goto et 641 FABP7 (fatty acid binding protein 7, brain) Godbout R, et al. with schizophrenia were up-regulated in the dorsolateral prefrontal cortex. Furthermore, single nucleotide polymorphism (SNP) analysis revealed an association between missense polymorphism Thr61Met 182C>T) and male patients with schizophrenia (Watanabe et al., 2007). In a separate study, FABP7 SNPs F704, F705 and F709 showed nominal association with bipolar disorder (Iwayama et al., 2010). Analysis of 6 FABP7 variants identified by polymorphic screen failed to identify any associations with autism or schizophrenia in 285 autistic and 1060 schizophrenic patients of Japanese descent (Maekawa et al., 2010). al., 2010). FABP7 immunostaining of 149 primary melanomas revealed an association between FABP7 expression and tumour thickness, as well as a trend towards increased relapse-free survival for patients who had tumors with low cytoplasmic FABP7 levels (Slipicevic et al., 2008). Knockdown of FABP7 in human melanoma cells resulted in decreased cell proliferation and invasion. There was no association between the nuclear expression of FABP7 and patient survival in this study (Slipicevic et al., 2008). Gene expression analysis of 87 primary melanomas and 68 metastatic melanoma, combined with immunohistochemical analysis of 37 paired primary and metastatic melanomas, showed significantly decreased FABP7 levels in metastatic melanoma compared to primary tumor tissue (Goto et al., 2010). In metastatic melanoma, FABP7 mRNA expression was associated with decreased relapsefree survival and overall survival (Goto et al., 2010). Loss of heterozygosity analysis using microsatellite markers specific to the FABP7 gene revealed that 10 of 20 metastatic melanomas (and 0 of 14 primary melanomas) had undergone loss of one FABP7 allele, leading the authors to postulate that genomic instability that favors loss of FABP7 expression may lead to better prognosis. References Feng L, Hatten ME, Heintz N. Brain lipid-binding protein (BLBP): a novel signaling system in the developing mammalian CNS. Neuron. 1994 Apr;12(4):895-908 Kurtz A, Zimmer A, Schnütgen F, Brüning G, Spener F, Müller T. The expression pattern of a novel gene encoding brain-fatty acid binding protein correlates with neuronal and glial cell development. Development. 1994 Sep;120(9):2637-49 Shi YE, Ni J, Xiao G, Liu YE, Fuchs A, Yu G, Su J, Cosgrove JM, Xing L, Zhang M, Li J, Aggarwal BB, Meager A, Gentz R. Antitumor activity of the novel human breast cancer growth inhibitor, mammary-derived growth inhibitor-related gene, MRG. Cancer Res. 1997 Aug 1;57(15):3084-91 Neurological disorders Note FABP7 is overexpressed in the brains of Down syndrome patients and has been postulated to contribute to Down syndrome-associated neurological disorders (Sánchez-Font et al., 2003). Pelsers et al. (2004) measured FABP7 levels in various parts of the adult human brain, with a range of 0,8 µg/g wet weight in the striatum and 3,1 µg/g in the frontal lobe. Measurement of FABP7 and FABP3 levels in the serum of patients with minor brain injuries identified both these FABPs as more sensitive at detecting brain injury than markers currently in use for this purpose. Similarly, serum FABP7 and FABP3 served as markers for individuals who had undergone ischaemic stroke (Wunderlich et al., 2005). FABP7 levels were also elevated in the serum of patients with neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease and other cognitive disorders. Although elevated levels of FABP7 were found in only one-third of patients, FABP7 is still the most discriminatory serum marker identified to date (Teunissen et al., 2011). The authors propose that elevated levels of FABP7 in serum may reflect damage to the central nervous system. FABP7-deficient mice have characteristics associated with schizophrenia such as decreased prepulse inhibition and shortened startle response latency (Watanabe et al., 2007). FABP7 RNA levels in the postmortem brains of male patients Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Godbout R, Bisgrove DA, Shkolny D, Day RS 3rd. Correlation of B-FABP and GFAP expression in malignant glioma. Oncogene. 1998 Apr 16;16(15):1955-62 Hohoff C, Spener F. Correspondence re: Y.E. Shi et al., Antitumor activity of the novel human breast cancer growth inhibitor, mammary-derived growth inhibitor-related gene, MRG. Cancer Res., 57: 3084-3091, 1997. 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A radial glia gene marker, fatty acid binding protein 7 (FABP7), is involved in proliferation and invasion of glioblastoma cells. PLoS One. 2012;7(12):e52113. doi: Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Godbout R, Poon HY, Liu RZ. FABP7 (fatty acid binding protein 7, brain). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):638-644. 644 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Gene Section Review GSTA1 (glutathione S-transferase alpha 1) Ana Savic-Radojevic, Tanja Radic Institute of Medical and Clinical Biochemistry, Faculty of Medicine, University of Belgrade, Serbia (ASR, TR) Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/GSTA1ID40764ch6p12.html DOI: 10.4267/2042/54029 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology Abstract genes (GSTA1, GSTA2, GSTA3, GSTA4, GSTA5) and seven pseudogenes (Morel et al., 2002). Review on GSTA1, with data on DNA/RNA, on the protein encoded and where the gene is implicated. Description The GSTA1 gene is approximately 12 kb in length and is closely flanked by other alpha class gene sequences. The complete sequence of the 1,7-kb intergenic region between exon 7 of an upstream pseudogene and exon 1 of the GSTA1 gene has been determined (Suzuki et al., 1993). Identity Other names: GST2, GSTA1-1, GTH1 HGNC (Hugo): GSTA1 Location: 6p12.2 Local order Between the LOC647169 (similar to glutathione transferase) and GSTA6P (glutathione S-transferase alpha 6 pseudogene) (according to PubMed). Note The GSTA1 gene is composed of 7 exons spanning a region of 12487 bases. Transcription The 1276-nucleotide transcript encodes a protein of 222 amino acid residues. Pseudogene An additional gene that encodes an uncharacterized Alpha class GST has been identified. The protein derived from this gene would have 19 amino acid substitutions compared with the GSTA1 isoenzyme. Several pseudogenes with single-base and/or complete exon deletions have been identified, but no reverse-transcribed pseudogenes have been detected (Suzuki et al., 1993). DNA/RNA Note The human alpha class genes are located in a cluster on chromosome 6p12 and comprise five functional GSTA1 gene. The GSTA1 gene spans a region of 12,5 kb composed of the seven exons (red) and six introns (green). Exons 1, 2, 3, 4, 5, 6 and 7 are 59 bp, 117 bp, 52 bp, 133 bp, 142 bp, 132 bp and 198 bp in length, respectively. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 645 GSTA1 (glutathione S-transferase alpha 1) Savic-Radojevic A, Radic T Crystal structure of human glutathione transferase (GST) A1-1 in complex with glutathione. Adapted from PDB (Grahn et al., 2006). Polymorphisms: GSTA1 has a functional three apparently linked single nucleotide polymorphisms (SNPs) in an SP1-responsive element within the proximal promoter (G-52A, C-69T and T-567G), plus at least four SNPs further upstream and a silent SNP A-375G. Two variants, GSTA1*A (-567T, 69C,-52G) and GSTA1*B (-67G, -69T, -52A), have been named according to the linked functional SNPs. Specifically, these substitutions result in differential expression with lower transcriptional activation of variant GSTA1*B than common GSTA1*A allele. It has been suggested that this genetic variation can change an individual's susceptibility to carcinogens and toxins, as well as, affect the efficacy of some drugs (Coles and Kadlubar, 2003). In addition, the linkage disequilibrium between GSTA1*A/GSTA1*B and GSTA2G335C (Ser112Thr) has been shown in Caucasians: specifically, GSTA1*A/GSTA2C335 (Thr112) and GSTA1*B/GSTA2G335 (Ser112) (Ning et al., 2004). It seems that the higher hepatic expression of GSTA1 enzyme in homozygous GSTA1 individuals is associated with the lower hepatic expression of GSTA2 in GSTA2C335 (Thr112) individuals (Coles et al., 2001a; Ning et al., 2004). Other haplotypes within this nomenclature but including SNPs C-115T, T-631G, and C-1142G also have been proposed (Bredschneider et al., 2002; Guy et al., 2004). Polymorphisms upstream of G-52C seem to have Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) little effect on GSTA1 expression (Morel et al., 2002). Protein Note Glutathione S-transferase A1 is N-terminally processed. Amino acids: 222. Calculated molecular mass: 25,63 kDa. Description The active GSTA1-1 enzyme is a homodimer, with each subunit containing a GSH-binding site (G-site) and a second adjacent hydrophobic binding site for the electrophilic substrate (H-site) (Wilce and Parker, 1994). The C-terminal region of GSTA1-1 contributes to the catalytic and noncatalytic ligand-binding functions of the enzyme, while the conserved G-site is located in the N-terminal domain (Balogh et al., 2009). Protein flexibility and dynamics in a molten globule-active site including the C-terminal α9 helix and the protruding ends of the α4-α5 helices result in achieving remarkable catalytic promiscuity of GSTA1-1 (Wu and Dong, 2012; Honaker et al., 2013). It has been proposed that the α9 helix may function as a mobile gate to the active-site cavity, controlling substrate access and product release. 646 GSTA1 (glutathione S-transferase alpha 1) Savic-Radojevic A, Radic T Structure determination and refinement of human alpha class glutathione transferase A1-1, and a comparison with the MU and PI class enzymes. Adapted from PDB (Sinning et al., 1993). Expression Function GSTA1-1 is highly expressed (as mRNA and protein) in liver, intestine, kidney, adrenal gland, pancreas and testis, while expression in a wide range of tissues is low (Hayes and Pulford, 1995; Coles et al., 2001a). Both positive and negative regulatory regions are present in the 5` noncoding region of GSTA1, including a polymorphic SP1binding site within the proximal promoter. Binding of the transcription factor AP1 has been suggested as a common mechanism for up-regulation of GSTs (Hayes and Pulford, 1995). The results of recent study also implied the role of a Kelch-like ECHassociated protein 1 (Keap1)-dependent signaling pathway for the induction of the constitutive GSTA1 expression during epithelial cell differentiation (Kusano et al., 2008). Regarding GSH-dependent ∆5-∆4 isomerase activity of this class of enzyme, it has been shown that steroidogenic factor 1 (SF-1) is involved in regulation of expression of GSTA genes (Matsumura et al., 2013). Aberrant overexpression has been observed in various malignancies such as colorectal (Hengstler et al., 1998) and lung cancer (Carmichael et al., 1988), while decrease in alpha class GSTs has been observed in stomach and liver tumors (Howie et al., 1990). A detailed recent review on GSTA1 can be found in Wu and Dong, 2012. Human GSTA1-1 enzyme catalyzes the GSHdependent detoxification of electrophiles showing highly promiscuous substrate selectivity for many structurally unrelated chemicals, including environmental carcinogens (e.g. benzo(a)pyrene diol epoxides), several alkylating chemotherapeutic agents (such as busulfan, chlorambucil, melphalan, phosphoramide mustard, cyclophosphamide, thiotepa), as well as, steroids and products of lipid degradation. GSTA1-1 is the most highly expressed GST of the liver and could therefore, be critical for "systemic" detoxification of electrophilic xenobiotics including carcinogens and drugs (Coles and Kadlubar, 2005). In addition to enzymatic detoxification, GSTA1 acts as modulator of mitogen-activated protein kinase (MAPK) signal transduction pathway via a mechanism involving protein-protein interactions. Namely, GSTA1 forms complexes with c-Jun Nterminal kinase (JNK), modifying JNK activation during cellular stress (Adnan et al., 2012). Thus, it is possible that GSTA1 confer drug resistance by two distinct means: by direct inactivation (detoxification) of chemotherapeutic drugs and by acting as inhibitors of MAPK pathway. Localisation The alpha class GSTs is showing strong intra-class sequence similarity (Balogh et al., 2009). Homology Cytosolic. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 647 GSTA1 (glutathione S-transferase alpha 1) Savic-Radojevic A, Radic T from acute myeloid leukemia patients, showing resistance to doxorubicin in vitro (Sargent et al., 1999). In addition, GSTA1 and CYP39A1 (member of cytochrome P450 family) polymorphisms were found to be associated with pharmacokinetics of busulfan, which is used in preparative regimens prior to stem cell transplantation in pediatric patients (ten Brink et al., 2013). Mutations Germinal None described so far. Somatic 36 mutations (COSMIC): 26 substitution-missense, 4 substitution-nonsense, 5 substitution-coding silent, 1 unknown type. Prostate cancer Note Genetic variants of GSTA1 and GSTT1 may modify prostate cancer risk, especially among smokers (Komiya et al., 2005). Implicated in Colorectal cancer Note Regarding the role of GSTA1 polymorphism in the risk of colorectal cancer, the results of epidemiological studies are still inconclusive. Several studies showed that GSTA1*B genotype (low hepatic expression) is associated with increased susceptibility to colorectal cancer, which imply the possible inefficient hepatic detoxification of food-derived carcinogen metabolite N-acetoxyPhIP (Coles et al., 2001b; Sweeney et al., 2002). In contrast, meta-analysis of Economopoulos representing the pooled analysis of four studies (1648 cases, 2039 controls) does not confer this association. Asthma Note Genetic alterations in GST enzymes may influence the detoxification of environmental toxic substances in airway and increase the risk of asthma. Thus, it has been shown that subjects with at least one allele -69T in the GSTA1 genotype have an increased risk of asthma (Polimanti et al., 2010). References Carmichael J, Forrester LM, Lewis AD, Hayes JD, Hayes PC, Wolf CR. Glutathione S-transferase isoenzymes and glutathione peroxidase activity in normal and tumour samples from human lung. Carcinogenesis. 1988 Sep;9(9):1617-21 Breast cancer Note The role of GSTA1 polymorphism in breast cancer risk was mainly based on investigation on response to chemotherapeutic drugs in these patients. In breast cancer patients on cyclophosphamide containing chemotherapy carriers of GSTA1*B/*B genotype showed significantly reduced five years risk of death in comparison to GSTA1*A homozygous carriers. This association was likely caused by decreased detoxification of the therapeutic metabolites of cyclophosphamide in GSTA1*B/*B patients (Sweeney et al., 2003). Howie AF, Forrester LM, Glancey MJ, Schlager JJ, Powis G, Beckett GJ, Hayes JD, Wolf CR. Glutathione Stransferase and glutathione peroxidase expression in normal and tumour human tissues. Carcinogenesis. 1990 Mar;11(3):451-8 Sinning I, Kleywegt GJ, Cowan SW, Reinemer P, Dirr HW, Huber R, Gilliland GL, Armstrong RN, Ji X, Board PG. Structure determination and refinement of human alpha class glutathione transferase A1-1, and a comparison with the Mu and Pi class enzymes. J Mol Biol. 1993 Jul 5;232(1):192-212 Suzuki T, Johnston PN, Board PG. Structure and organization of the human alpha class glutathione Stransferase genes and related pseudogenes. Genomics. 1993 Dec;18(3):680-6 Bladder cancer Note Recent investigation indicates that the GSTA1-low activity genotype in combination with the GSTM1null genotype significantly increases the risk of bladder cancer in smokers (Matic et al., 2013). In addition, it seems that GSTA1 polymorphism may influence vulnerability to oxidative DNA damage, thereby contributing to the malignant potential of transitional cell carcinoma (Savic-Radojevic et al., 2013). Wilce MC, Parker MW. Structure and function of glutathione S-transferases. Biochim Biophys Acta. 1994 Mar 16;1205(1):1-18 Hayes JD, Pulford DJ. The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance. Crit Rev Biochem Mol Biol. 1995;30(6):445-600 Hengstler JG, Böttger T, Tanner B, Dietrich B, Henrich M, Knapstein PG, Junginger T, Oesch F. Resistance factors in colon cancer tissue and the adjacent normal colon tissue: glutathione S-transferases alpha and pi, glutathione and aldehyde dehydrogenase. Cancer Lett. 1998 Jun 5;128(1):105-12 Myeloid leukemia Note Aberant overexpression of both GSTA1 and GSTA2 proteins was found in blast cells derived Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Sargent JM, Williamson C, Hall AG, Elgie AW, Taylor CG. Evidence for the involvement of the glutathione pathway in 648 GSTA1 (glutathione S-transferase alpha 1) Savic-Radojevic A, Radic T drug resistance in AML. Adv Exp Med Biol. 1999;457:2059 Biol Crystallogr. 2006 Feb;62(Pt 2):197-207 Kusano Y, Horie S, Shibata T, Satsu H, Shimizu M, Hitomi E, Nishida M, Kurose H, Itoh K, Kobayashi A, Yamamoto M, Uchida K. Keap1 regulates the constitutive expression of GST A1 during differentiation of Caco-2 cells. Biochemistry. 2008 Jun 10;47(23):6169-77 Coles BF, Morel F, Rauch C, Huber WW, Yang M, Teitel CH, Green B, Lang NP, Kadlubar FF. Effect of polymorphism in the human glutathione S-transferase A1 promoter on hepatic GSTA1 and GSTA2 expression. Pharmacogenetics. 2001a Nov;11(8):663-9 Balogh LM, Le Trong I, Kripps KA, Tars K, Stenkamp RE, Mannervik B, Atkins WM. Structural analysis of a glutathione transferase A1-1 mutant tailored for high catalytic efficiency with toxic alkenals. Biochemistry. 2009 Aug 18;48(32):7698-704 Coles B, Nowell SA, MacLeod SL, Sweeney C, Lang NP, Kadlubar FF. The role of human glutathione S-transferases (hGSTs) in the detoxification of the food-derived carcinogen metabolite N-acetoxy-PhIP, and the effect of a polymorphism in hGSTA1 on colorectal cancer risk. Mutat Res. 2001b Oct 1;482(1-2):3-10 Economopoulos KP, Sergentanis TN. GSTM1, GSTT1, GSTP1, GSTA1 and colorectal cancer risk: a comprehensive meta-analysis. Eur J Cancer. 2010 Jun;46(9):1617-31 Bredschneider M, Klein K, Mürdter TE, Marx C, Eichelbaum M, Nüssler AK, Neuhaus P, Zanger UM, Schwab M. Genetic polymorphisms of glutathione Stransferase A1, the major glutathione S-transferase in human liver: consequences for enzyme expression and busulfan conjugation. Clin Pharmacol Ther. 2002 Jun;71(6):479-87 Polimanti R, Piacentini S, Moscatelli B, Pellicciotti L, Manfellotto D, Fuciarelli M. GSTA1, GSTO1 and GSTO2 gene polymorphisms in Italian asthma patients. Clin Exp Pharmacol Physiol. 2010 Aug;37(8):870-2 Adnan H, Antenos M, Kirby GM. The effect of menadione on glutathione S-transferase A1 (GSTA1): c-Jun Nterminal kinase (JNK) complex dissociation in human colonic adenocarcinoma Caco-2 cells. Toxicol Lett. 2012 Oct 2;214(1):53-62 Morel F, Rauch C, Coles B, Le Ferrec E, Guillouzo A. The human glutathione transferase alpha locus: genomic organization of the gene cluster and functional characterization of the genetic polymorphism in the hGSTA1 promoter. Pharmacogenetics. 2002 Jun;12(4):277-86 Wu B, Dong D. Human cytosolic glutathione transferases: structure, function, and drug discovery. Trends Pharmacol Sci. 2012 Dec;33(12):656-68 Sweeney C, Coles BF, Nowell S, Lang NP, Kadlubar FF. Novel markers of susceptibility to carcinogens in diet: associations with colorectal cancer. Toxicology. 2002 Dec 27;181-182:83-7 Honaker MT, Acchione M, Zhang W, Mannervik B, Atkins WM. Enzymatic detoxication, conformational selection, and the role of molten globule active sites. J Biol Chem. 2013 Jun 21;288(25):18599-611 Coles BF, Kadlubar FF. Detoxification of electrophilic compounds by glutathione S-transferase catalysis: determinants of individual response to chemical carcinogens and chemotherapeutic drugs? Biofactors. 2003;17(1-4):115-30 Matic M, Pekmezovic T, Djukic T, Mimic-Oka J, Dragicevic D, Krivic B, Suvakov S, Savic-Radojevic A, PljesaErcegovac M, Tulic C, Coric V, Simic T. GSTA1, GSTM1, GSTP1, and GSTT1 polymorphisms and susceptibility to smoking-related bladder cancer: a case-control study. Urol Oncol. 2013 Oct;31(7):1184-92 Sweeney C, Ambrosone CB, Joseph L, Stone A, Hutchins LF, Kadlubar FF, Coles BF. Association between a glutathione S-transferase A1 promoter polymorphism and survival after breast cancer treatment. Int J Cancer. 2003 Mar 1;103(6):810-4 Matsumura T, Imamichi Y, Mizutani T, Ju Y, Yazawa T, Kawabe S, Kanno M, Ayabe T, Katsumata N, Fukami M, Inatani M, Akagi Y, Umezawa A, Ogata T, Miyamoto K. Human glutathione S-transferase A (GSTA) family genes are regulated by steroidogenic factor 1 (SF-1) and are involved in steroidogenesis. FASEB J. 2013 Aug;27(8):3198-208 Guy CA, Hoogendoorn B, Smith SK, Coleman S, O'Donovan MC, Buckland PR. Promoter polymorphisms in glutathione-S-transferase genes affect transcription. Pharmacogenetics. 2004 Jan;14(1):45-51 Ning B, Wang C, Morel F, Nowell S, Ratnasinghe DL, Carter W, Kadlubar FF, Coles B. Human glutathione Stransferase A2 polymorphisms: variant expression, distribution in prostate cancer cases/controls and a novel form. Pharmacogenetics. 2004 Jan;14(1):35-44 Savic-Radojevic A, Djukic T, Simic T, Pljesa-Ercegovac M, Dragicevic D, Pekmezovic T, Cekerevac M, Santric V, Matic M. GSTM1-null and GSTA1-low activity genotypes are associated with enhanced oxidative damage in bladder cancer. Redox Rep. 2013;18(1):1-7 Coles BF, Kadlubar FF. Human alpha class glutathione Stransferases: genetic polymorphism, expression, and susceptibility to disease. Methods Enzymol. 2005;401:9-42 ten Brink MH, van Bavel T, Swen JJ, van der Straaten T, Bredius RG, Lankester AC, Zwaveling J, Guchelaar HJ. Effect of genetic variants GSTA1 and CYP39A1 and age on busulfan clearance in pediatric patients undergoing hematopoietic stem cell transplantation. Pharmacogenomics. 2013 Nov;14(14):1683-90 Komiya Y, Tsukino H, Nakao H, Kuroda Y, Imai H, Katoh T. Human glutathione S-transferase A1, T1, M1, and P1 polymorphisms and susceptibility to prostate cancer in the Japanese population. J Cancer Res Clin Oncol. 2005 Apr;131(4):238-42 This article should be referenced as such: Grahn E, Novotny M, Jakobsson E, Gustafsson A, Grehn L, Olin B, Madsen D, Wahlberg M, Mannervik B, Kleywegt GJ. New crystal structures of human glutathione transferase A1-1 shed light on glutathione binding and the conformation of the C-terminal helix. Acta Crystallogr D Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Savic-Radojevic A, Radic T. GSTA1 (glutathione Stransferase alpha 1). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):645-649. 649 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Gene Section Short Communication MOAP1 (Modulator Of Apoptosis 1) Gamze Ayaz, Mesut Muyan Department of Biological Sciences, Middle East Technical University, Ankara, Turkey (GA, MM) Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/MOAP1ID46494ch14q32.html DOI: 10.4267/2042/54030 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology Abstract Protein Short communication on MOAP1, with data on DNA/RNA, on the protein encoded and where the gene is implicated. Note MOAP1 is a short-lived protein with a half-life of 25 minutes. It is degraded by the ubiquitin-proteosome system (Fu et al., 2007). Identity Description Other names: MAP-1, PNMA4 HGNC (Hugo): MOAP1 Location: 14q32.12 MOAP-1 is a 351 amino-acid long protein with a molecular mass of 39.5 kDa. Isoelectric point (pI) of MOAP-1 is 4.939 at pH 7.0. MOAP-1 contains a BH3 (Bcl-2 homology 3) like domain required for homodimerization and interaction with Bcl-2 associated X (Bax) protein. Under normal condition, MOAP1 is held as an inactive conformation through intramolecular interactions. Interaction between RASSF1A (rasassociation domain family 1, isoform A) and MOAP1 reduces the inhibitory intramolecular interaction of MOAP1 and allows MOAP1 through the BH3 like domain to bind Bax (Baksh et al., 2005). DNA/RNA Description Human MOAP1 gene contains three exons and the encoding sequence of 1056 bases is in the exon 3. Transcription MOAP1 is a 351 amino-acid long protein. Pseudogene No reported pseudogenes. The MOAP1 gene is located in the reverse strand. Exons are shown as boxes and introns as lines. The filled box in the exon 3 is the coding sequence of MOAP1. Numbers below represent the size of exon or intron in base pair. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 650 MOAP1 (Modulator Of Apoptosis 1) Ayaz G, Muyan M the downregulation of MOAP1 expression and the aggressiveness of breast cancer (Law, 2012). Expression MOAP1 is expressed in the adipose, adrenal, blood, brain, breast, colon and heart. MOAP1 is expressed in higher level in the heart and brain. To be noted Note miRNA: It is reported that miR-1228 prevents cellular apoptosis by binding to the 3'UTR of MOAP1 mRNA, thereby decreasing MOAP-1 protein levels (Yan and Zhao, 2012). Localisation MOAP-1 protein localizes in the cytoplasm (Law, 2012) and also seen in the mitochondria (Tan et al., 2005). Function References MOAP1 is a BH3-like protein that acts as a proapoptotic molecule (Tan et al., 2001). When overexpressed, MOAP1 induces caspase-dependent apoptosis in mammalian cells (Tan et al., 2005). Studies showed that TNFα stimulation recruits RASSF1A and MOAP1 to death receptors complexes. This recruitment leads to the association of RASSF1A with MOAP1 and to the induction of a conformational change in MOAP1. This results in the opening of the BH3 domain to allow MOAP1 to interact with Bax. Bax is subsequently inserted into the mitochondrial membrane leading to apoptosis (Baksh et al., 2005; Foley et al., 2008). Tan KO, Tan KM, Chan SL, Yee KS, Bevort M, Ang KC, Yu VC. MAP-1, a novel proapoptotic protein containing a BH3-like motif that associates with Bax through its Bcl-2 homology domains. J Biol Chem. 2001 Jan 26;276(4):2802-7 Baksh S, Tommasi S, Fenton S, Yu VC, Martins LM, Pfeifer GP, Latif F, Downward J, Neel BG. The tumor suppressor RASSF1A and MAP-1 link death receptor signaling to Bax conformational change and cell death. Mol Cell. 2005 Jun 10;18(6):637-50 Tan KO, Fu NY, Sukumaran SK, Chan SL, Kang JH, Poon KL, Chen BS, Yu VC. MAP-1 is a mitochondrial effector of Bax. Proc Natl Acad Sci U S A. 2005 Oct 11;102(41):14623-8 Fu NY, Sukumaran SK, Yu VC. Inhibition of ubiquitinmediated degradation of MOAP-1 by apoptotic stimuli promotes Bax function in mitochondria. Proc Natl Acad Sci U S A. 2007 Jun 12;104(24):10051-6 Homology Also highly conserved in rat (Rattus norvegicus) and mouse (Mus musculus), human MOAP1 protein shares 99% amino acids identity with that of chimpanzee (Pan troglodytes) (Law et al., 2012). Foley CJ, Freedman H, Choo SL, Onyskiw C, Fu NY, Yu VC, Tuszynski J, Pratt JC, Baksh S. Dynamics of RASSF1A/MOAP-1 association with death receptors. Mol Cell Biol. 2008 Jul;28(14):4520-35 Mutations Law J.. MOAP1: A Candidate Tumor Suppressor Protein Master Thesis, University of Alberta, Canada, 2012 http://hdl.handle.net/10402/era.24828. Note Gene mutations have not been described yet. Law J, Yu VC, Baksh S.. Modulator of Apoptosis 1: A Highly Regulated RASSF1A-Interacting BH3-Like Protein. Mol Biol Int. 2012;2012:536802. doi: 10.1155/2012/536802. Epub 2012 Jun 14. Implicated in Breast cancer Yan B, Zhao JL.. miR-1228 prevents cellular apoptosis through targeting of MOAP1 protein. Apoptosis. 2012 Jul;17(7):717-24. doi: 10.1007/s10495-012-0710-9. Disease Microarray analysis of 176 primary, treatmentnaive breast cancer and of 10 normal breast tissue samples suggests that the MOAP1 gene expression is significantly down-regulated in breast cancer. It appears that there is a positive correlation between Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) This article should be referenced as such: Ayaz G, Muyan M. MOAP1 (Modulator Of Apoptosis 1). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):650651. 651 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Gene Section Short Communication PHLDA1 (pleckstrin homology-like domain, family A, member 1) Maria Aparecida Nagai Discipline of Oncology, Department of Radiology and Oncology, Medical School, University of Sao Paulo, Center for Translational Investigation in Oncology, Cancer Institute from Sao Paulo State, Sao Paulo, Brazil (MAN) Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/PHLDA1ID41707ch12q15.html DOI: 10.4267/2042/54031 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology PHLDA1 protein has a modular structure containing a central pleckstrin homology-like domain (PHL) and prolin-glutamine (PQ) and proline-histidine (PH) repeats in the C-terminal region (see figure above). Abstract Short communication on PHLDA1, with data on DNA/RNA, on the protein encoded and where the gene is implicated. Expression Identity DNA/RNA PHLDA1 is widely expressed in mammalian tissues displaying cytoplasmic, vesicle membrane, plasma membrane and nuclear subcellular localization. PHLDA1 expression is up-regulated by estrogen, IGF-1 (insulin-like growth factor 1), FGF (fibroblast growth factor), TPA (phorbol ester), and ER (endoplamic reticulum)-stress agents such as homocysteine, tunicamicyne, and farnesol. Description Localisation PHLDA1 gene contains 2 exons, 1 intervening sequence and spans 6,3 kb of genomic DNA. Cytoplasm, vesicle membrane, plasma membrane, nucleus. Transcription Function 1,2 kb mRNA. Protein binding. Several evidences have implicate PHLDA1 as a potential transcriptional activator that acts as a pro-apoptotic and antiproliferative factor, however the mechanisms by which PHLDA1 mediates cell survival is still under investigation. Other names: DT1P1B11, PHRIP, TDAG51 HGNC (Hugo): PHLDA1 Location: 12q21.2 Local order: Minus strand. Pseudogene Not identified. Protein Homology Description PHLDA2 (pleckstrin homology-like domain, family A, member 2) and PHLDA3 (pleckstrin homologylike domain, family A, member 3) are paralogs for PHLDA1. The PHLDA1 gene encodes for a 401 amino acid protein that is a member of the evolutionarily conserved pleckstrin homology-like domain family. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 652 PHLDA1 (pleckstrin homology-like domain, family A, member 1) Nagai MA Schematic representation of the modular structure of PHLDA1 protein. PHL: pleckstrin homology-like domain spanning amino acids residues from 150 to 283; QQ: proline/glutamine rich sequence (aa residues from 189 to 204); PQ: prolineglutamine tracts (aa residues from 311 to 346); PH: proline-histidine-rich tracts (aa residues from 352 to 389); *: indicates phosphorylation sites. significantly better in patients with tumors that were negative for PHLDA1, and a multivariate analysis suggested that PHLDA1 is an independent prognostic factor in OSCC patients (CoutinhoCamillo et al., 2013). Mutations Note Short genetic variation - dbSNP: rs139162669, rs73385441, rs74620794, rs147230079, rs76437300, rs186978611, rs140610935, rs144470255, rs79545253, rs147644129. Colon cancer Note Altered PHLDA1 expression has been shown to be associated with the process of intestinal tumorigenesis (Sakthianandeswaren et al., 2011). Implicated in Melanoma Note PHLDA1 expression was associated with reduced cell growth and colony formation and with increased apoptotic rates and drug sensitivity in melanoma cell lines. Loss of PHLDA1 has been correlated with melanoma progression (Neef et al., 2002). Basal cell carcinoma Breast cancer Atherosclerosis Note Down-regulation of PHLDA1 mRNA and protein expression is frequently observed in primary invasive breast tumours. Down-regulation of PHLDA1 protein has been shown to be a strong predictor of poor prognosis for breast cancer patients, indicating that reduced PHLDA1 expression contribute for breast cancer progression and might serve as useful prognostic biomarker of disease outcome (Nagai et al., 2007). Note In vivo and in vitro studies demonstrated that increased PHLDA1 expression induced by homocysteine promotes detachment-mediated programmed cell death and contributes to the development of atherosclerosis (Hossain et al., 2003). Genetic variant in an intergenic region of the PHLDA1 gene (rs2367446) has been shown to be associated with the development of cardiovascular diseases (Hossain et al., 2013). Note PHLDA1 has also been shown to be a follicular and epithelial stem cell marker (Ohyama et al., 2006; Sakthianandeswaren et al., 2011) with potential to differentiates between trichoepithelioma and basal cell carcinoma (Sellheyer and Nelson, 2011). Oral cancer Epilepsy Note Reduced expression of PHLDA1 was observed in 60,7% of oral squamous cell carcinomas (OSCC), especially in well-differentiated tumors. Positive PHLDA1 immunostaining was associated with advanced clinical stages of the disease, suggesting that PHLDA1 has a functional role in oral tumorigenesis. Overall and disease-free survival rates were Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Note PHLDA1 expression has been shown to be higher in the anterior temporal neocortex from patients with intractable epilepsy when compared with the levels observed in the neocortex from the control group, suggesting a possible association of PHLDA1 in the physiopathology of the disease (Xi et al., 2007). 653 PHLDA1 (pleckstrin homology-like domain, family A, member 1)Nagai MA References neocortex of patients with intractable epilepsy. Neurosci Lett. 2007 Sep 20;425(1):53-8 Park CG, Lee SY, Kandala G, Lee SY, Choi Y. A novel gene product that couples TCR signaling to Fas(CD95) expression in activation-induced cell death. Immunity. 1996 Jun;4(6):583-91 Marchiori AC, Casolari DA, Nagai MA. Transcriptional upregulation of PHLDA1 by 17beta-estradiol in MCF-7 breast cancer cells. Braz J Med Biol Res. 2008 Jul;41(7):579-82 Johnson EO, Chang KH, de Pablo Y, Ghosh S, Mehta R, Badve S, Shah K. PHLDA1 is a crucial negative regulator and effector of Aurora A kinase in breast cancer. J Cell Sci. 2011 Aug 15;124(Pt 16):2711-22 Frank D, Mendelsohn CL, Ciccone E, Svensson K, Ohlsson R, Tycko B. A novel pleckstrin homology-related gene family defined by Ipl/Tssc3, TDAG51, and Tih1: tissue-specific expression, chromosomal location, and parental imprinting. Mamm Genome. 1999 Dec;10(12):1150-9 Sakthianandeswaren A, Christie M, D'Andreti C, Tsui C, Jorissen RN, Li S, Fleming NI, Gibbs P, Lipton L, Malaterre J, Ramsay RG, Phesse TJ, Ernst M, Jeffery RE, Poulsom R, Leedham SJ, Segditsas S, Tomlinson IP, Bernhard OK, Simpson RJ, Walker F, Faux MC, Church N, Catimel B, Flanagan DJ, Vincan E, Sieber OM. PHLDA1 expression marks the putative epithelial stem cells and contributes to intestinal tumorigenesis. Cancer Res. 2011 May 15;71(10):3709-19 Kuske MD, Johnson JP. Assignment of the human PHLDA1 gene to chromosome 12q15 by radiation hybrid mapping. Cytogenet Cell Genet. 2000;89(1-2):1 Neef R, Kuske MA, Pröls E, Johnson JP. Identification of the human PHLDA1/TDAG51 gene: down-regulation in metastatic melanoma contributes to apoptosis resistance and growth deregulation. Cancer Res. 2002 Oct 15;62(20):5920-9 Sellheyer K, Nelson P. Follicular stem cell marker PHLDA1 (TDAG51) is superior to cytokeratin-20 in differentiating between trichoepithelioma and basal cell carcinoma in small biopsy specimens. J Cutan Pathol. 2011 Jul;38(7):542-50 Hossain GS, van Thienen JV, Werstuck GH, Zhou J, Sood SK, Dickhout JG, de Koning AB, Tang D, Wu D, Falk E, Poddar R, Jacobsen DW, Zhang K, Kaufman RJ, Austin RC. TDAG51 is induced by homocysteine, promotes detachment-mediated programmed cell death, and contributes to the cevelopment of atherosclerosis in hyperhomocysteinemia. J Biol Chem. 2003 Aug 8;278(32):30317-27 Nagai MA.. PHLDA1 (pleckstrin homology-like domain, family A, member). Encyclopedia of Signaling Molecules 2012, pp 1365 - 1369. ISBN: 978-4419-04060-7 (Editor: Sandun Choi) Coutinho-Camillo CM, Lourenco SV, Nonogaki S, Vartanian JG, Nagai MA, Kowalski LP, Soares FA.. Expression of PAR-4 and PHLDA1 is prognostic for overall and disease-free survival in oral squamous cell carcinomas. Virchows Arch. 2013 Jul;463(1):31-9. doi: 10.1007/s00428-013-1438-9. Epub 2013 Jun 9. Oberg HH, Sipos B, Kalthoff H, Janssen O, Kabelitz D. Regulation of T-cell death-associated gene 51 (TDAG51) expression in human T-cells. Cell Death Differ. 2004 Jun;11(6):674-84 Toyoshima Y, Karas M, Yakar S, Dupont J, Lee Helman, LeRoith D. TDAG51 mediates the effects of insulin-like growth factor I (IGF-I) on cell survival. J Biol Chem. 2004 Jun 11;279(24):25898-904 Hossain GS, Lynn EG, Maclean KN, Zhou J, Dickhout JG, Lhotak S, Trigatti B, Capone J, Rho J, Tang D, McCulloch CA, Al-Bondokji I, Malloy MJ, Pullinger CR, Kane JP, Li Y, Shiffman D, Austin RC.. Deficiency of TDAG51 protects against atherosclerosis by modulating apoptosis, cholesterol efflux, and peroxiredoxin-1 expression. J Am Heart Assoc. 2013 May 17;2(3):e000134. doi: 10.1161/JAHA.113.000134. Ohyama M, Terunuma A, Tock CL, Radonovich MF, PiseMasison CA, Hopping SB, Brady JN, Udey MC, Vogel JC. Characterization and isolation of stem cell-enriched human hair follicle bulge cells. J Clin Invest. 2006 Jan;116(1):24960 This article should be referenced as such: Nagai MA, Fregnani JH, Netto MM, Brentani MM, Soares FA. Down-regulation of PHLDA1 gene expression is associated with breast cancer progression. Breast Cancer Res Treat. 2007 Nov;106(1):49-56 Nagai MA. PHLDA1 (pleckstrin homology-like domain, family A, member 1). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):652-654. Xi ZQ, Wang LY, Sun JJ, Liu XZ, Zhu X, Xiao F, Guan LF, Li JM, Wang L, Wang XF. TDAG51 in the anterior temporal Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 654 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Gene Section Review ADAMTS15 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif, 15) Santiago Cal, Alvaro J Obaya Departamento de Bioquimica y Biologia Molecular, Instituto Universitario de Oncologia (IUOPA), Universidad de Oviedo, 33006, Asturias, Spain (SC), Biologia Funcional, Instituto Universitario de Oncologia (IUOPA), Universidad de Oviedo, 33006, Asturias, Spain (AJO) Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/ADAMTS15ID45587ch11q24.html DOI: 10.4267/2042/54032 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology 8 exons, spans approximately 27.66 Kb of genomic DNA in the centromere-to-telomere orientation. The translation initiation codon is located to exon 1, and the stop codon to exon 8. protein, with an estimated molecular weight of 103,2 kDa. ADAMTS-15 shares a structural multidomain complex architecture with the rest of the members of the ADAMTS family. This organization includes a signal peptide, a prodomain involved in maintaining enzyme latency and a catalytic domain that contains the consensus sequence HEXXHGXXHD involved in the coordination of the zinc atom necessary for catalytic activiy of the enzyme. This sequence ends in an Asp residue which distinguishes ADAMTSs from other metalloproteases such as MMPs. Following this catalytic region there are several other domains characterized as disintegrin-like domain, a central thrombospondin-1 (TSP-1) motif, a cysteine-rich domain, a spacer region and two more TSP-1 domains (Cal et al., 2002). Protein Expression Abstract Review on ADAMTS15, with data on DNA/RNA, on the protein encoded and where the gene is implicated. Identity HGNC (Hugo): ADAMTS15 Location: 11q24.3 DNA/RNA Description ADAMTS15 cDNA was originally cloned from both, a human liver and kidney fetal cDNA library (Cal et al., 2002). Description The open reading fame encodes a 950 amino acid Domain organization of ADAMTS-15. Pro: prodomain; TSP: thrombospondin type-1 domains. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 655 ADAMTS15 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif, 15) Later on, in the search for proteinases and proteinase inhibitors in articular cartilage from femoral heads of patients with end-stage osteoarthritis (OA) Kevorkian et al. found high levels of ADAMTS-15 expression in samples from both, OA patients as well as normal controls (Kevorkian et al., 2004). In relation with ADAMTS-15 participation in tumor progression its expression has been described in either normal cells or cells adjacent or marginal to cancer tissue in samples from colon adenocarcinoma as well as in samples from head and neck squamous cell carcinoma (Viloria et al., 2009; Stokes et al., 2010). Additionally ADAMTS-15 presence has also been detected in some breast and prostate cancer cell lines (Molokwu et al., 2010). chicken (Refseq: XM_417874), and zebrafish (Refseq: XM_001341842). Mutations Somatic ADAMTS15 was identified as one of the so-called CAN genes found to be mutated in a small set of colorectal cancers (Sjöblom et al., 2006). Two heterozigous somatic mutations were described out of eleven human cancer samples (cDNA: 2309A>G, cDNA: 2632T>G). Functional relevance of mutations found in colorectal cancer were described for a deleterious single base mutation 24544∆G affecting the two carboxy-terminal thrombospondin motifs of ADAMTS-15 (Viloria et al., 2009). The derived truncated form of ADAMTS-15 (ADAMTS15_G849fs) is barely found in the pericellular space of the cell being mostly liberated to the culture media. Functional studies revealed ADAMTS15_G849fs not showing the anti-tumoral properties of full length ADAMTS-15. In the same study, three other mutations where identified, a base pair mutation affecting the second TSP-1 domain (24616C>T), a silent base pair change (13777C>T) and another base deletion generating a completely truncated form of ADAMTS-15 (366∆) (Viloria et al., 2009). Localisation Extracellular, mostly pericellular. Function Few studies describe ADAMTS-15 function beyond those describing its participation in cancer and osteoartritic processes. Regarding cancer, ADAMTS-15 has recently emerged as a putative tumor suppresor gene since it is downregulated in breast cancer, and functionally inactivated through specific mutations in colorectal cancer (Porter et al., 2004; Porter et al., 2006; Viloria et al., 2009). In addition, aberrant expression of ADAMTS-15 is implicated in prostate cancer progression (Molokwu et al., 2010). The latest apparently results from the relationship between ADAMTS-15 expression and versican degradation. Thus, ADAMTS-15 seems to be acting as a versicandegrading enzyme whose accumulation potentially contributes to prostate cancer pathology (Cross et al., 2005). In this regard, versican seems to be one of the targets of ADAMTS-15 proteolityc activity which involves this protein in processes such as cancer or skeletal muscle fiber formation (Croos et al., 2005; Stupka et al., 2013; Dancevic et al., 2013). Implicated in Various cancers Note ADAMTS-15 has recently emerged as a putative tumor suppresor gene since it is downregulated in breast cancer, and functionally inactivated through specific mutations in colorectal cancer (Porter et al., 2006; López-Otín et al., 2009; Viloria et al., 2009). In addition, aberrant expression of ADAMTS-15 is implicated in prostate cancer progression (Cross et al., 2005; Molokwu et al., 2010). The first indication regarding a potential protective role for ADAMTS15 derived from the observation that low ADAMTS15 expression levels coupled to high ADAMTS8 levels conferred poor prognosis to breast cancer patients (Porter et al., 2006). Moreover, ADAMTS15 was identified as one of the so-called CAN genes found to be mutated in a small set of colorectal cancers (Sjöblom et al., 2006). Functional support to the putative relevance of ADAMTS-15 as a tumor suppresor protease was described after finding four additional mutations in ADAMTS-15 gene sequence in human colon carcinomas (Viloria et al., 2009). Two of the new mutations resulted in the generation of truncated forms of ADAMTS-15, one of them lacking the last two thrombospondin domains whereas the other originating a complete ADAMTS-15 knock-down. Homology ADAMTS-15 belongs to the A Disintegrin And Metalloprotease Domains with ThromboSpondin motifs (ADAMTS) family, which consists of 19 secreted zinc metalloproteinases (Porter et al., 2005). All members of the family share the same structural domain design. ADAMTS-15 is, among all the members, closely related to ADAMTS-1 which suggested its involvement in angiogenic processes (Cal et al., 2002). The ADAMTS15 gene is conserved in chimpanzee (Refseq: XM_522253), macaque (Refseq: XM_001113698), dog (Refseq: XM_005620295), cow (Refseq: NM_001192390), mouse (Refseq: NM_001024139), rat (Refseq: NM_001106810), Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Cal S, Obaya AJ 656 ADAMTS15 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif, 15) Cal S, Obaya AJ Functional analysis revealed that the presence of the two last thrombospondin domains is important for the pericellular loacalization of ADAMTS-15 and affects the anti-tumoral function of full length ADAMTS-15 (Viloria et al., 2009; Dancevic et al., 2013). More recently, ADAMTS-15 has been described as a head and neck squamous cell carcinoma (HNSCC)-associated proteinase since its expression is elevated (together with ADAMTS-1 and ADAMTS-8) in areas surrounding HNSCC tumor microenvironment (Demircan et al., 2009; Stokes et al., 2010). In addition, these three members of the ADAMTS family have elevated expression levels in HNSCC tumor versus normal tissue and in HNSCC derived cell lines vs normal keratinocytes (Stokes et al., 2010). ADAMTS-15 has also been indirectly involved in androgen-mediated prostate cancer growth and proliferation, function that depends on ADAMTS15 versicanolytic activity (Cross et al., 2005; Molokwu et al., 2010). Molokwu et al identified one androgen-responsive element (ARE) in ADAMTS-15 promoter and 12 more AREs in its gene sequence. In the same article the authors demonstrated ADAMTS-15 reduction both, at mRNA and protein levels, in the presence of dihidrotestorone (DHT). ADAMTS-15 down-regulation in prostate cancer resulted in high versican levels which is a poor prognosis indicator in these type of tumors (Ricciardelli et al., 1998; Luo et al., 2002; Molokwu et al., 2010). Breast cancer Colon cancer Porter S, Clark IM, Kevorkian L, Edwards DR. The ADAMTS metalloproteinases. Biochem J. 2005 Feb 15;386(Pt 1):15-27 Note ADAMTS15 elevated expression correlates with favorable outcome in patients with breast cancer (Porter et al., 2006). References Ricciardelli C, Mayne K, Sykes PJ, Raymond WA, McCaul K, Marshall VR, Horsfall DJ. Elevated levels of versican but not decorin predict disease progression in early-stage prostate cancer. Clin Cancer Res. 1998 Apr;4(4):963-71 Cal S, Obaya AJ, Llamazares M, Garabaya C, Quesada V, López-Otín C. Cloning, expression analysis, and structural characterization of seven novel human ADAMTSs, a family of metalloproteinases with disintegrin and thrombospondin1 domains. Gene. 2002 Jan 23;283(1-2):49-62 Luo J, Dunn T, Ewing C, Sauvageot J, Chen Y, Trent J, Isaacs W. Gene expression signature of benign prostatic hyperplasia revealed by cDNA microarray analysis. Prostate. 2002 May 15;51(3):189-200 Kevorkian L, Young DA, Darrah C, Donell ST, Shepstone L, Porter S, Brockbank SM, Edwards DR, Parker AE, Clark IM. Expression profiling of metalloproteinases and their inhibitors in cartilage. Arthritis Rheum. 2004 Jan;50(1):13141 Porter S, Scott SD, Sassoon EM, Williams MR, Jones JL, Girling AC, Ball RY, Edwards DR. Dysregulated expression of adamalysin-thrombospondin genes in human breast carcinoma. Clin Cancer Res. 2004 Apr 1;10(7):2429-40 Cross NA, Chandrasekharan S, Jokonya N, Fowles A, Hamdy FC, Buttle DJ, Eaton CL. The expression and regulation of ADAMTS-1, -4, -5, -9, and -15, and TIMP-3 by TGFbeta1 in prostate cells: relevance to the accumulation of versican. Prostate. 2005 May 15;63(3):269-75 Note ADAMTS15 expression inversely correlates with histopathologic differentiation grade in human colorectal carcinomas when analyzing ADAMTS15 inmunostaining in normal colon epithelia, welldifferentiated tumors, moderately differentiated tumors, and poorly differentiated colorectal carcinomas (Viloria et al., 2009). Porter S, Span PN, Sweep FC, Tjan-Heijnen VC, Pennington CJ, Pedersen TX, Johnsen M, Lund LR, Rømer J, Edwards DR. ADAMTS8 and ADAMTS15 expression predicts survival in human breast carcinoma. Int J Cancer. 2006 Mar 1;118(5):1241-7 Sjöblom T, Jones S, Wood LD, Parsons DW, Lin J, Barber TD, Mandelker D, Leary RJ, Ptak J, Silliman N, Szabo S, Buckhaults P, Farrell C, Meeh P, Markowitz SD, Willis J, Dawson D, Willson JK, Gazdar AF, Hartigan J, Wu L, Liu C, Parmigiani G, Park BH, Bachman KE, Papadopoulos N, Vogelstein B, Kinzler KW, Velculescu VE. The consensus coding sequences of human breast and colorectal cancers. Science. 2006 Oct 13;314(5797):268-74 Head and neck squamous carcinoma (HNSCC) Note ADAMTS15 mRNA levels, together with those of other ADAMTS members (ADAMTS1, ADAMTS4, ADAMTS5, ADAMTS8, ADAMTS9), were reduced in HNSCC primary tumors compared with paired non-cancerous tissues (Demircan et al., 2009). Regarding tumor microenvironment ADAMTS15 expression is elevated in adjacent and margin tissue when compared with tumor center tissue (Stokes et al., 2010). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Demircan K, Gunduz E, Gunduz M, Beder LB, Hirohata S, Nagatsuka H, Cengiz B, Cilek MZ, Yamanaka N, Shimizu K, Ninomiya Y. Increased mRNA expression of ADAMTS metalloproteinases in metastatic foci of head and neck cancer. Head Neck. 2009 Jun;31(6):793-801 López-Otín C, Palavalli LH, Samuels Y. Protective roles of matrix metalloproteinases: from mouse models to human cancer. Cell Cycle. 2009 Nov 15;8(22):3657-62 Viloria CG, Obaya AJ, Moncada-Pazos A, Llamazares M, Astudillo A, Capellá G, Cal S, López-Otín C. Genetic 657 ADAMTS15 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif, 15) inactivation of ADAMTS15 metalloprotease in human colorectal cancer. Cancer Res. 2009 Jun 1;69(11):4926-34 disintegrin-like and metalloproteinase domain with thrombospondin-1 repeats-15: a novel versican-cleaving proteoglycanase. J Biol Chem. 2013 Dec 27;288(52):37267-76 Molokwu CN, Adeniji OO, Chandrasekharan S, Hamdy FC, Buttle DJ. Androgen regulates ADAMTS15 gene expression in prostate cancer cells. Cancer Invest. 2010 Aug;28(7):698-710 Stupka N, Kintakas C, White JD, Fraser FW, Hanciu M, Aramaki-Hattori N, Martin S, Coles C, Collier F, Ward AC, Apte SS, McCulloch DR. Versican processing by a disintegrin-like and metalloproteinase domain with thrombospondin-1 repeats proteinases-5 and -15 facilitates myoblast fusion. J Biol Chem. 2013 Jan 18;288(3):1907-17 Stokes A, Joutsa J, Ala-Aho R, Pitchers M, Pennington CJ, Martin C, Premachandra DJ, Okada Y, Peltonen J, Grénman R, James HA, Edwards DR, Kähäri VM. Expression profiles and clinical correlations of degradome components in the tumor microenvironment of head and neck squamous cell carcinoma. Clin Cancer Res. 2010 Apr 1;16(7):2022-35 This article should be referenced as such: Cal S, Obaya AJ. ADAMTS15 (ADAM Metallopeptidase With Thrombospondin Type 1 Motif, 15). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):655-658. Dancevic CM, Fraser FW, Smith AD, Stupka N, Ward AC, McCulloch DR. Biosynthesis and expression of a Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Cal S, Obaya AJ 658 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Gene Section Review ADRB2 (adrenoceptor beta 2, surface) Denise Tostes Oliveira, Diego Mauricio Bravo-Calderón Department of Stomatology, Area of Pathology, Bauru School of Dentistry - University of Sao Paulo, Bauru, Brazil (DTO, DMBC) Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/ADRB2ID43818ch5q32.html DOI: 10.4267/2042/54033 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology 2006). The receptor is comprised of 413 amino acid residues of approximately 46500 daltons (Johnson, 2006). β2 adrenergic receptor is N-glycosylated at amino acids 6, 15, and 187; these are important for roper insertion of the receptor into the membrane as well as for agonist trafficking (McGraw and Liggett, 2005; Johnson, 2006). Abstract Review on ADRB2, with data on DNA/RNA, on the protein encoded and where the gene is implicated. Identity Expression Other names: ADRB2R, ADRBR, B2AR, BAR, BETA2AR HGNC (Hugo): ADRB2 Location: 5q32 β2 adrenergic receptor is widely distributed, this protein is expressed by airway smooth muscle (3040000 per cell), epithelial and endothelial cells of the lung, smooth muscle of blood vessels, skeletal muscle, mast cells, lymphocytes, oral and skin keratinocytes and also by diverse cancer cells (Kohm and Sanders, 2001; Lutgendorf et al., 2003; Johnson, 2006; Sood et al., 2006; Thaker et al., 2006; Yang et al., 2006; Sastry et al., 2007; Yu et al., 2007; Liu et al., 2008a; Liu et al., 2008b; Shang et al., 2009; Sivamani et al., 2009; Yang et al., 2009; Bernabé et al., 2011; Bravo-Calderón et al., 2011-2012; Steenhuis et al., 2011; Zhang et al., 2011; Loenneke et al., 2012). DNA/RNA Description ADBR2 gene spans about 2,04 kb and consists of one exon. Transcription ADBR2 no has introns in either their coding or untranslated sequences. The primary transcripts are processed at their 5' and 3' ends like other premessenger RNAs, but no splicing is needed. Localisation Pseudogene β2 adrenergic receptor is a transmembrane protein. Like all GPCRs, the β2 adrenergic receptor has seven transmembrane a domains that form a pocket containing binding sites for agonists and competitive antagonists (McGraw and Liggett, 2005; Johnson, 2006). There are 3 extracellular loops, with one being the amino terminus, and 3 intracellular loops, with a carboxy terminus (McGraw and Liggett, 2005; Johnson, 2006). No pseudogenes have been reported. Protein Description β2 adrenergic receptor is a member of the superfamily of G-protein coupled receptors (GPCRs) (McGraw and Liggett, 2005; Johnson, Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 659 ADRB2 (adrenoceptor beta 2, surface) Oliveira DT, Bravo-Calderón DM Activation of protein kinase A (PKA) by signal transduction of β2 adrenergic receptor (adapted of Rosenbaum et al., 2009). foradrenergic receptors in these experimental effects. Norepinephrine was later found to increase the in vitro invasive potential of ovarian cancer cells, an effect that was blocked by propranolol (Sood et al., 2006). Norepinephrine also increased tumor cell expression of matrix metalloproteinase-2 (MMP-2) and MMP-9, and pharmacologic blockade of MMPs abrogated the effects of norepinephrine on tumor cell invasive potential (Sood et al., 2006). In the same way, Thaker et al. (Thaker et al., 2006) correlated chronic behavioral stress with higher levels of tissue catecholamines and more invasive growth of ovarian carcinoma cells in an orthotopic mouse model. These effects were mediated through β2 adrenergic receptor activation of PKA signaling pathway (Thaker et al., 2006). Tumors in stressed animals showed increased vascularization and enhanced expression of VEGF, MMP2 and MMP9; these effects could be abrogated by propranolol (Thaker et al., 2006). Function Agonist binding of β2 adrenergic receptor results in activation of Gs protein. The Gs protein a subunit stimulates adenylyl cyclase to generate cyclic 3'-5'adenosine monophosphate (cAMP), which in sequence activates the cAMP-dependent protein kinase A (PKA) and the agonist-occupied receptor is phosphorylated. After phosphorylation, the receptor switches its coupling specificity to Gi. GTP-bound Giα dissociates from the heterodimeric Gβγ, and free Gβγ subunits mediate activation of the MAP kinase signaling pathway in the same way as Gi-coupled receptors. Increase of intracellular cAMP levels leads diverse cell functions as cell proliferation, differentiation, angiogenesis and migration (Daaka et al., 1997). Implicated in Ovarian carcinoma Prostate cancer Note Reverse transcriptase-PCR studies indicated constitutive expression of β2 adrenergic receptor on ovarian carcinoma cell lines (Lutgendorf et al., 2003). Lutgendorf et al. (Lutgendorf et al., 2003) investigated the effects of norepinephrine and isoproterenol (a nonspecific-adrenergic agonist) on the production of vascular endothelial growth factor (VEGF) by ovarian cancer cell lines; and found that both, norepinephrine and isoproterenol, significantly enhanced VEGF production. These effects were blocked by thenon-specific β antagonist propranolol, supporting a role Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Note β2 adrenergic receptor signaling was related to prostate cancer cell progression (Sastry et al., 2007; Zhang et al., 2011). β2 adrenergic receptor activation of PKA signaling pathway has been associated with reduction of sensitivity of prostate cancer cells to apoptosis (Sastry et al., 2007) and promotion of cell proliferation and cell migration (Zhang at al., 2011). Contrastingly, other investigation demonstrated that the genetic silencing of β2 adrenergic receptor increases cell migration and invasion of normal 660 ADRB2 (adrenoceptor beta 2, surface) Oliveira DT, Bravo-Calderón DM prostate cells and that the weak expression of this protein is associated with metastases and with worst survival rates in prostate cancer patients (Yu et al., 2007). carcinoma cells lines (Yang et al., 2006); as well upregulated the production of VEGF, interleukin (IL)-8, and IL-6 in human melanoma tumor cell lines (Yang et al., 2009). Esophageal squamous cell carcinoma References Daaka Y, Luttrell LM, Lefkowitz RJ. Switching of the coupling of the beta2-adrenergic receptor to different G proteins by protein kinase A. Nature. 1997 Nov 6;390(6655):88-91 Note Liu et al. (Liu et al., 2008b) demonstrated that stimulation of β2 adrenergic receptor with epinephrine significantly increase the esophageal cancer cell proliferation accompanied by elevation of the expression of VEGF, VEGF receptor VEGFR-1 and VEGFR-2. In addition, it has been shown that the epidermal growth factor mediates the mitogenic signals in esophageal cancer cells through transactivation of β2 adrenergic receptor (Liu et al., 2008a). Kohm AP, Sanders VM. Norepinephrine and beta 2adrenergic receptor stimulation regulate CD4+ T and B lymphocyte function in vitro and in vivo. Pharmacol Rev. 2001 Dec;53(4):487-525 Lutgendorf SK, Cole S, Costanzo E, Bradley S, Coffin J, Jabbari S, Rainwater K, Ritchie JM, Yang M, Sood AK. Stress-related mediators stimulate vascular endothelial growth factor secretion by two ovarian cancer cell lines. Clin Cancer Res. 2003 Oct 1;9(12):4514-21 Oral squamous cell carcinoma (OSCC) McGraw DW, Liggett SB. Molecular mechanisms of beta2adrenergic receptor function and regulation. Proc Am Thorac Soc. 2005;2(4):292-6; discussion 311-2 Note Genetic and protein expression of β2 adrenergic receptor was demonstrated in OSCC by using RTPCR assay, Western blot and immunohistochemistry (Shang et al., 2009; Bernabé et al., 2011; Bravo-Calderón et al., 2011-2012). Investigations performed in different oral cancer cell lines demonstrated that β2 adrenergic receptor signaling by norepinephrine increases cell proliferation and invasion, and upregulates interleukin-6 (IL-6) gene expression and protein release (Shang et al., 2009; Bernabé et al., 2011). Furthermore, Shang et al. (Shang et al., 2009) reported that malignant cell positive immunoexpression of β2-AR was significantly correlated with age, tumor size, clinical stage and cervical lymph node metastasis in OSCC patients, and that β2-AR may play an important role in the formation and metastasis of oral cancer. However, a retrospective clinical study of a large number of patients showed that patients with OSCC who exhibited strong β2-AR immunohistochemical expression by malignant epithelial cells demonstrated higher survival rates compared to patients with weak/negative β2-AR expression (Bravo-Calderón et al., 2011-2012). Therefore, further clinical and laboratory studies are warranted to elucidate the role of β2 adrenergic receptor activation in oral squamous cell carcinoma. Johnson M. Molecular mechanisms of beta(2)-adrenergic receptor function, response, and regulation. J Allergy Clin Immunol. 2006 Jan;117(1):18-24; quiz 25 Sood AK, Bhatty R, Kamat AA, Landen CN, Han L, Thaker PH, Li Y, Gershenson DM, Lutgendorf S, Cole SW. Stress hormone-mediated invasion of ovarian cancer cells. Clin Cancer Res. 2006 Jan 15;12(2):369-75 Thaker PH, Han LY, Kamat AA, Arevalo JM, Takahashi R, Lu C, Jennings NB, Armaiz-Pena G, Bankson JA, Ravoori M, Merritt WM, Lin YG, Mangala LS, Kim TJ, Coleman RL, Landen CN, Li Y, Felix E, Sanguino AM, Newman RA, Lloyd M, Gershenson DM, Kundra V, Lopez-Berestein G, Lutgendorf SK, Cole SW, Sood AK. Chronic stress promotes tumor growth and angiogenesis in a mouse model of ovarian carcinoma. Nat Med. 2006 Aug;12(8):939-44 Yang EV, Sood AK, Chen M, Li Y, Eubank TD, Marsh CB, Jewell S, Flavahan NA, Morrison C, Yeh PE, Lemeshow S, Glaser R. Norepinephrine up-regulates the expression of vascular endothelial growth factor, matrix metalloproteinase (MMP)-2, and MMP-9 in nasopharyngeal carcinoma tumor cells. Cancer Res. 2006 Nov 1;66(21):10357-64 Sastry KS, Karpova Y, Prokopovich S, Smith AJ, Essau B, Gersappe A, Carson JP, Weber MJ, Register TC, Chen YQ, Penn RB, Kulik G. Epinephrine protects cancer cells from apoptosis via activation of cAMP-dependent protein kinase and BAD phosphorylation. J Biol Chem. 2007 May 11;282(19):14094-100 Yu J, Cao Q, Mehra R, Laxman B, Yu J, Tomlins SA, Creighton CJ, Dhanasekaran SM, Shen R, Chen G, Morris DS, Marquez VE, Shah RB, Ghosh D, Varambally S, Chinnaiyan AM. Integrative genomics analysis reveals silencing of beta-adrenergic signaling by polycomb in prostate cancer. Cancer Cell. 2007 Nov;12(5):419-31 Various cancers Note β2 adrenergic receptor was also immunohistochemically identified in nasopharyngeal carcinoma (Yang et al., 2006) and in melanoma (Yang et al., 2009). Norepinephrine treatment increased MMP-2, MMP-9, and VEGF levels in culture supernatants of nasopharyngeal Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Liu X, Wu WK, Yu L, Li ZJ, Sung JJ, Zhang ST, Cho CH. Epidermal growth factor-induced esophageal cancer cell proliferation requires transactivation of betaadrenoceptors. J Pharmacol Exp Ther. 2008a Jul;326(1):69-75 Liu X, Wu WK, Yu L, Sung JJ, Srivastava G, Zhang ST, Cho CH. Epinephrine stimulates esophageal squamous- 661 ADRB2 (adrenoceptor beta 2, surface) Oliveira DT, Bravo-Calderón DM cell carcinoma cell proliferation via beta-adrenoceptordependent transactivation of extracellular signal-regulated kinase/cyclooxygenase-2 pathway. J Cell Biochem. 2008b Sep 1;105(1):53-60 carcinoma cells. Brain Behav Immun. 2011 Mar;25(3):57483 Bravo-Calderón DM, Oliveira DT, Marana AN, Nonogaki S, Carvalho AL, Kowalski LP. Prognostic significance of beta2 adrenergic receptor in oral squamous cell carcinoma. Cancer Biomark. 2011-2012;10(1):51-9 Rosenbaum DM, Rasmussen SG, Kobilka BK. The structure and function of G-protein-coupled receptors. Nature. 2009 May 21;459(7245):356-63 Steenhuis P, Huntley RE, Gurenko Z, Yin L, Dale BA, Fazel N, Isseroff RR. Adrenergic signaling in human oral keratinocytes and wound repair. J Dent Res. 2011 Feb;90(2):186-92 Shang ZJ, Liu K, Liang de F. Expression of beta2adrenergic receptor in oral squamous cell carcinoma. J Oral Pathol Med. 2009 Apr;38(4):371-6 Zhang P, He X, Tan J, Zhou X, Zou L. β-arrestin2 mediates β-2 adrenergic receptor signaling inducing prostate cancer cell progression. Oncol Rep. 2011 Dec;26(6):1471-7 Sivamani RK, Pullar CE, Manabat-Hidalgo CG, Rocke DM, Carlsen RC, Greenhalgh DG, Isseroff RR. Stress-mediated increases in systemic and local epinephrine impair skin wound healing: potential new indication for beta blockers. PLoS Med. 2009 Jan 13;6(1):e12 Loenneke JP, Wilson JM, Thiebaud RS, Abe T, Lowery RP, Bemben MG. β2 Adrenoceptor signaling-induced muscle hypertrophy from blood flow restriction: is there evidence? Horm Metab Res. 2012 Jun;44(7):489-93 Yang EV, Kim SJ, Donovan EL, Chen M, Gross AC, Webster Marketon JI, Barsky SH, Glaser R. Norepinephrine upregulates VEGF, IL-8, and IL-6 expression in human melanoma tumor cell lines: implications for stress-related enhancement of tumor progression. Brain Behav Immun. 2009 Feb;23(2):267-75 This article should be referenced as such: Oliveira DT, Bravo-Calderón DM. ADRB2 (adrenoceptor beta 2, surface). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):659-662. Bernabé DG, Tamae AC, Biasoli ÉR, Oliveira SH. Stress hormones increase cell proliferation and regulates interleukin-6 secretion in human oral squamous cell Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 662 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Gene Section Review IRF4 (interferon regulatory factor 4) Vipul Shukla, Runqing Lu Department of Genetics, Cell Biology & Anatomy, University of Nebraska Medical Center, Omaha, NE 68118, USA (VS, RL) Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/IRF4ID231ch6p25.html DOI: 10.4267/2042/54034 This article is an update of : Rasi S, Gaidano G. IRF4 (interferon regulatory factor 4). Atlas Genet Cytogenet Oncol Haematol 2009;13(12):941-943. This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology Abstract mRNA is expressed at high levels in lymphoid tissues, in skin and in tonsils. Review on IRF4, with data on DNA/RNA, on the protein encoded and where the gene is implicated. Protein Identity Description Protein length: 451 amino acids. Calculated molecular weight of 51,8 kDa. Other names: IRF-4, LSIRF, MUM1, NF-EM5 HGNC (Hugo): IRF4 Location: 6p25.3 Local order: IRF4 is located on chromosome 6 at the telomeric extremity of the short arm, and lies between the DUSP22 (dual specificity phosphatase 22) and EXOC2 (exocyst complex component 2) genes. Note IRF4 belongs to the IRF (interferon regulatory factors) family of transcription factors and is a critical transcriptional regulator of immune system development and function. Expression IRF4 protein is expressed predominantly in blood cells. However, its expression can also be detected in adipocytes and melanocytes. In blood cells, expression of IRF4 can be detected in T, B, DC and macrophages. Expression of IRF4 in T and B cells is strongly induced by antigen receptor signaling. Localisation Nucleus. Function In the immune system, IRF4 is critical for development and maturation of multiple lineages of blood cells. In T cells development, IRF4 is essential for the differentiation of Th1, Th2, Th9, Th17 and T reg subsets. In B lymphocytes, IRF4 promotes light chain rearrangement and transcription and is critical for B cell development at the pre-B stage. IRF4 antagonizes Notch signaling and limits the size of marginal zone B cells (Simonetti et al., 2013). DNA/RNA Description Gene of 19,4 kb with 9 exons and 8 introns. Exon 1, the 5' part of exon 2 and the 3' part of exon 9 are non coding. Transcription Length of the transcript is 5314 bp. Coding sequence: CDS 114-1469. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 663 IRF4 (interferon regulatory factor 4) Shukla V, Lu R In addition, IRF4 is essential for class-switching and plasma cell differentiation. In B cells, IRF4 interacts with Ets family of trancription factor (PU.1/spi-B) through EICE site whereas in T cells, IRF4 interacts with AP-1 family of trancription factor (BATF) through AICE site. Also, IRF4 is required for the differentiation of dendritic cells (DCs), particularly the CD11b(+) subset (Schlitzer et al., 2013). In macrophages, IRF4 promotes the differentiation and polarisation to the M2 subtype also known as the tumor associated macrophages. Recent studies have identified a role of IRF4 in adipocyte biology. IRF4 has been shown to regulate enzymes required for lipolysis in adipocytes. Therefore, an adipocyte specific deletion of IRF4 causes enhanced lipid synthesis, dysregulated lipid homeostasis eventually leading to obesity. Interestingly, in melanocytes IRF4 was recently identified to cooperate with another transcription factor, MITF to positively regulate the expression of tyrosinase gene required for melanin synthesis. Additionally, the SNPs in the IRF4 gene locus have been identified as risk alleles for developing melanoma. course. IRF4 is obligatory required for the terminal differetiation of mature B cells to plasma cells and has been shown to play a central role in the pathogenesis of MM. IRF4 is recurrently translocated and juxtaposed to the IgH promoter t(6;14)(p25;q32) in a significant proportion (~21%) of MM cases. More commonly, IRF4 have been shown to be overexpressed without genetic alterations in majority of MM cases and MM cells are particularly sensitive to the down-regulation of IRF4. Cytogenetics t(6;14)(p25;q32) --> IRF4 - IgH. Hybrid/Mutated gene The translocation juxtaposes the IgH locus to the IRF4 gene. Oncogenesis The precise mechanism for pathogenesis of MM in presence of high levels of IRF4 is mediated by an autoregulatory loop established between IRF4 and c-myc in MM cells. Recently, IRF4 has been shown to regulate caspase-10 leading to disruption of normal autophagy mechanisms in MM cells thereby, causing prolonged survival of these cells. Homology Chronic lymphocytic leukemia (CLL) Among IRF family members, IRF4 is highly homologous to IRF8. Disease CLL is the most common adult leukemia in the western countries. It is a heterogeneous B-cell malignancy marked by progressive accumulation of CD5 positive mature B lymphocytes. A Genome Wide Association Study (GWAS) recently identified SNPs in the 3' UTR of IRF4 gene locus in patients with CLL. The individuals carrying the risk alleles harboring the SNPs have lower levels of IRF4 and poorer outcomes compared to individuals carrying the non-risk allele. Another study identified mutations in the DNA binding domain of IRF4 in a small subset (1,5%) of CLL cases. More recently, using two distinct murine genetic models, it has been shown that low levels of IRF4 are causally related to the development of CLL. Prognosis CLL patients with low levels of IRF4 have aggressive disease course and poor prognosis. Mutations Germinal SNPs in the IRF4 gene locus have been identified in patients with chronic lymphocytic leukemia and melanoma. Somatic Somatic mutations in DNA binding domain of IRF4 have been identified in a small subset (1,5%) of chronic lymphocytic leukemia (CLL) patients. Implicated in Multiple myeloma (MM) Disease Multiple myeloma (MM) is a plasma cell derived malignancy with a particularly aggressive clinical Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 664 IRF4 (interferon regulatory factor 4) Shukla V, Lu R Hodgkins lymphoma (HL) Cytogenetics Although reciprocal translocations are extremely rare in CLL, a translocation disrupting IRF4 gene locus t(1;6)(p35.3;p25.2) was identified in a small subset of CLL patients with aggressive disease. Hybrid/Mutated gene Mutations in the DNA binding domain of IRF4 with a yet undefined function in B cells were identified in a small subset of CLL cases. Oncogenesis The precise mechanism for oncogenesis of CLL in presence of low levels of IRF4 is not yet known. Disease Hodgkins lymphoma (HL) is an enigmatic B cell malignancy that is characterized by lack of expression of several B cell markers. The Hodgkin and Reed Sternberg (HRS) cells present in HL cases are presumably derived from germinal center B cells. IRF4 is overexpressed in majority of classical HL cases and is shown to mediate the survival of these cells. Paradoxically, the SNPs in IRF4 linked to its lower expression levels and associated with the development of CLL are also shown to be linked to the risk of developing HL. Oncogenesis Whether the overexpression of IRF4 in HRS cells of HL is causal is unclear. However, some studies have linked the survival and proliferation of HRS cells to the expression of IRF4. Diffused large B cell lymphoma (DLBCL) Disease Diffuse large B cell lymphoma represents a heterogeneous malignancy that arises spontaneously or develop from pre-existing leukemia. On the basis of gene expression profiling DLBCL is divided into three distinct subtypes namely the germinal center subtype (GCB), the activated B cell subtype (ABC) and the mediastinal subtype. The three subtypes presumably arise from three distinct B cell subtypes. IRF4 is primarily overexpressed in the ABC type of DLBCL while GCB subtype is marked by lower expression of IRF4. Prognosis IRF4 is overexpressed in the ABC type DLBCL which is most aggressive form of DLBCL and have poorer patient outcomes compared to other subtypes. Cytogenetics IRF4 is overexpressed in a small group of patients with a reciprocal translocation between IgG locus and the IRF4 t(1;6)(p35.3;p25.2). The patients carrying the translocation primarily belong to GCB or follicular lymphoma grade 3 type is associated with favorable patient outcomes. Oncogenesis IRF4 induces the expression of transcription factor Blimp-1 and directly suppresses the expression of Bcl-6 to allow terminal differentiation of activated B cells to plasma cells. However, this molecular network is short circuited in ABC DLBCL by recurrent mutational inactivation of Blimp-1. Additionally, mutations located in the promoter region of Bcl-6 that disrupt the IRF4 binding sites and leads to enhanced expression of Bcl-6 were identified in a small group of patients. These genetic events disrupt the molecular network required for plasma cell differentiation. However, the precise functional role of IRF4 in pathogenesis of ABC type DLBCL is not well defined. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Primary cutaneous anaplastic large cell lymphoma (C-ALCL) Disease Primary cutaneous anaplastic large cell lymphoma (C-ALCL) is a T cell lymphoma with an indolent disease course and presence of tumor lesions in the skin. The lesions in C-ALCL almost never spread extra-cutaneously and often regress spontaneously. IRF4 is overexpressed in C-ALCL but not in the more aggressive form of the disease known as peripheral T cell lymphoma not otherwise specified (PTCL-NOS). The overexpression of IRF4 in some cases is associated with a recurrent translocations a subset of them placing the IRF4 gene next to the T cell receptor alpha (TCRA) promoter t(6;14)(p25;q11.2). Other translocations identified do not involve TCRA. Cytogenetics IRF4 is translocated primarily in the C-ALCL however the precise breakpoints are not defined. In a small subset of the cases with translocations IRF4 is juxtaposed to the TCRA locus t(6;14)(p25;q11.2). B cell acute lymphoblastic leukemia (B-ALL) Disease B cell acute lymphoblastic leukemia (B-ALL) is a B cell malignancy derived from early B cells. IRF4 is shown to play a tumor suppressive role in BALL. IRF4 is shown to suppress the oncogenesis of both BCR-ABL and c-myc induced B-ALL. Oncogenesis IRF4 inhibits B-ALL by regulating the expression of negative regulators of cell cycle p27. 665 IRF4 (interferon regulatory factor 4) Shukla V, Lu R the screen map to a putative enhancer region in the IRF4 gene locus. Oncogenesis Recently, the SNP identified in IRF4 locus were demonstrated to decrease IRF4 expression by disruption of specific transcription factor binding sites. Additionally, IRF4 corroborates with micropthalmia associated transcription factor (MITF) to regulate the expression of enzyme tyrosinase responsible for melanin production. These studies point towards a critical role for IRF4 in melanocyte biology and also its association with skin cancer. Chronic myeloid leukemia (CML) Disease Chronic myeloid leukemia (CML) is a myeloproliferative disorder marked by clonal expansion of granulocytes. It is associated with a hallmark translocation and presence of a fusion BCR-ABL protein in majority of patients. IRF4 is shown to be underexpressed in CML patients along with its highly homologous family member IRF8. However the functional role of IRF4 in CML is not well characterized. Virus implicated malignancies Disease Viruses like Epstein Barr virus (EBV), human T cell leukemia virus-1 (HTLV1) and Kaposi Sarcoma associated herpes virus (KSHV/HHV-8) are implicated in B cell malignancies, adult T cell leukemia (ATL) and primary effusion lymphoma (PEL) respectively. The proteins encoded by these viruses, directly or indirectly activate NF-kB signaling which in turn activates the expression of IRF4. As a result IRF4 is overexpressed in these virus implicated malignancies. The knockdown of IRF4 in EBV transformed B cells lead to downregulation of genes involved in cellular proliferation. The role of IRF4 in HTLV-1 induced ATL is not clear however few reports indicate its involvement in regulation of cell cycle associated genes. The role of IRF4 in KSHV induced kaposi's sarcoma and PEL is ambiguous. KSHV encodes viral homologs of cellular IRFs called vIRFs. The vIRF4 is shown to inhibit the function of cellular IRF4 leading to induction of lytic cycle for KSHV replication. Oncogenesis The role of IRF4 in these viral implicated malignancies is still unclear. However, the activation status of NF-kB by these viruses invariably co-relates with IRF4 expression in these cells. References Grossman A, Mittrücker HW, Nicholl J, Suzuki A, Chung S, Antonio L, Suggs S, Sutherland GR, Siderovski DP, Mak TW. Cloning of human lymphocyte-specific interferon regulatory factor (hLSIRF/hIRF4) and mapping of the gene to 6p23-p25. Genomics. 1996 Oct 15;37(2):229-33 Iida S, Rao PH, Butler M, Corradini P, Boccadoro M, Klein B, Chaganti RS, Dalla-Favera R. Deregulation of MUM1/IRF4 by chromosomal translocation in multiple myeloma. Nat Genet. 1997 Oct;17(2):226-30 Mittrücker HW, Matsuyama T, Grossman A, Kündig TM, Potter J, Shahinian A, Wakeham A, Patterson B, Ohashi PS, Mak TW. Requirement for the transcription factor LSIRF/IRF4 for mature B and T lymphocyte function. Science. 1997 Jan 24;275(5299):540-3 Tsuboi K, Iida S, Inagaki H, Kato M, Hayami Y, Hanamura I, Miura K, Harada S, Kikuchi M, Komatsu H, Banno S, Wakita A, Nakamura S, Eimoto T, Ueda R. MUM1/IRF4 expression as a frequent event in mature lymphoid malignancies. Leukemia. 2000 Mar;14(3):449-56 Chang CC, Lorek J, Sabath DE, Li Y, Chitambar CR, Logan B, Kampalath B, Cleveland RP. Expression of MUM1/IRF4 correlates with clinical outcome in patients with B-cell chronic lymphocytic leukemia. Blood. 2002 Dec 15;100(13):4671-5 Falini B, Mason DY. Proteins encoded by genes involved in chromosomal alterations in lymphoma and leukemia: clinical value of their detection by immunocytochemistry. Blood. 2002 Jan 15;99(2):409-26 Ito M, Iida S, Inagaki H, Tsuboi K, Komatsu H, Yamaguchi M, Nakamura N, Suzuki R, Seto M, Nakamura S, Morishima Y, Ueda R. MUM1/IRF4 expression is an unfavorable prognostic factor in B-cell chronic lymphocytic leukemia (CLL)/small lymphocytic lymphoma (SLL). Jpn J Cancer Res. 2002 Jun;93(6):685-94 Skin cancer Disease Skin cancer is associated with malignant or nonmalignant lesions on the skin. Based on the cell of origin, skin cancer can be divided into three types: basal cell carcinoma, squamous cell carcinoma and melanoma. The differential skin pigmentation induced by melanin production alters the risk for skin cancer. Particularly individuals with light skin tones and hence low melanin secretion are more predisposed to developing skin cancer. Until recently there were no known reports for a role of IRF4 in melanocytes. However recently, SNPs identified in the IRF4 gene locus have been shown to be associated with skin pigmentation and the risk for developing skin cancer. The SNP identified in Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Mamane Y, Grandvaux N, Hernandez E, Sharma S, Innocente SA, Lee JM, Azimi N, Lin R, Hiscott J. Repression of IRF-4 target genes in human T cell leukemia virus-1 infection. Oncogene. 2002 Oct 3;21(44):6751-65 Nishiya N, Yamamoto K, Imaizumi Y, Kohno T, Matsuyama T. Identification of a novel GC-rich binding protein that binds to an indispensable element for constitutive IRF-4 promoter activity in B cells. Mol Immunol. 2004 Jul;41(9):855-61 Honma K, Udono H, Kohno T, Yamamoto K, Ogawa A, Takemori T, Kumatori A, Suzuki S, Matsuyama T, Yui K. Interferon regulatory factor 4 negatively regulates the production of proinflammatory cytokines by macrophages 666 IRF4 (interferon regulatory factor 4) Shukla V, Lu R in response to LPS. Proc Natl Acad Sci U S A. 2005 Nov 1;102(44):16001-6 Oct;11(10):936-44 Eguchi J, Wang X, Yu S, Kershaw EE, Chiu PC, Dushay J, Estall JL, Klein U, Maratos-Flier E, Rosen ED. 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IRF4 (interferon regulatory factor 4). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):663-667. 667 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Gene Section Review PLCG1 (Phospholipase C, Gamma 1) Rebeca Manso Pathology Department, Fundacion Conchita Rabago, IIS "Fundacion Jimenez Diaz", E-28040 Madrid, Spain (RM) Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/PLCG1ID44163ch20q12.html DOI: 10.4267/2042/54035 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology Expression Abstract PLCG1 is expressed ubiquitously, especially in the brain, thymus, intestine and lungs. Additionally, PLCG1 is overexpressed in numerous cancer types such as human colorectal cancer (Noh et al., 1994), breast carcinoma (Arteaga et al., 1991), prostate carcinoma (Peak et al., 2008), familial adenomatous polyposis (Park et al., 1994) and human skins under hypeproliferative conditions (Nanney et al., 1992). Review on PLCG1, with data on DNA/RNA, on the protein encoded and where the gene is implicated. Identity Other names: NCKAP3, PLC-II, PLC1, PLC148, PLCgamma1 HGNC (Hugo): PLCG1 Location: 20q12 Local order: From the cytosol. Localisation PLCG1 localizes predominantly in the plasmatic membrane, cytoplasm and nucleus. DNA/RNA Function Description PLCG1 is a protein involved in multiple cellular processes. A potent inhibitor of PLCG1 (U-73122) has been reported to inhibit PLCG1-dependent processes in cells (Smith et al., 1990; Thompson et al., 1991; Thomas et al., 2003; Li et al., 2005). The inhibition of PLCG1 may be an important mechanism for an antiproliferative effect on the human cancer cells. Role in the production of the second messenger molecules: PLCG1 mediates the production of diacylglycerol (DAG) and inositol 1,4,5trisphosphate (IP3) from the hydrolysis of phosphatidynositol-4,5-bisphosphate (PIP2) (Williams et al., 1996). These second messengers are essential for T cell activation (Lin et al., 2001). The PLCG1 gene spans 38.762 kb on the genomic DNA. The gene includes 32 exons. Transcription There are two transcript variants: 5205 bp (isoform a) and 5202 bp (isoform b). Protein Description The PLCG1 protein encodes two alternative isoforms: variant a (P19174-1)-1290 amino acids, 148.53 Da; variant b (P19174-2)-1291 amino acids, 148.66 Da. Figure 1. Schematic diagram of PLCG1 location on chromosome 20. PLCG1 localizes to chromosome 20q12, which is represented graphically. PLCG1 gene spans 38.762 kb on the genomic DNA. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 668 PLCG1 (Phospholipase C, Gamma 1) Manso R Figure 2. Schematic representation of the domains of PLCG1. The protein contains eight domains, four of which are unique to PLCG family. The PLCG 'specific array' of domains, comprising a "split" PH domain flanking two tandem SH2 domains and one SH3 domain, is inserted between the two halves (X and Y) of the TIM-barrel catalytic domain. Several other domains including two PH domains, one C2 domain and one EF hand motifs. The numering of the amino acid residues is for human PLCG1 (Suh et al., 2008; Bunney and Katan, 2011). Role in cellular proliferation: PLCG1 is associated with tumor development, and it is overexpressed in some tumors (Shin et al., 2007). This overexpression stimulates MMP-3 expression. PLCG1 is required for metastasis development (Sala et al., 2008). Role in angiogenesis: PLCG1 plays an important role in angiogenesis (Husain et al., 2010). PLCG1 is activated by vascular endothelial growth factor receptor-2 (VEGFR-2) in endothelial cells (Singh et al., 2007) and in neoplastic Barrett's cells (Zhang et al., 2013). Role in the regulation of intracellular signaling: PLCG1 plays a role in mediating T-cell activities downstream of TCR activity. PLCG1 can be activated by receptor tyrosine kinases: EGFR (Nishibe et al., 1990; Wu et al., 2009), PDGFR (Larose et al., 1993), FGFR (Peters et al., 1992), NGFR (Middlemas et al., 1994) and HGFR (Davies et al., 2008). PLCG1 is a molecule associate with lipid rafts, it translocates from the cytosol to lipid rafts during TCR signaling (Verí et al., 2001). Role in the mobilization of Ca2+: this process is to activate phosphatase calcineurin, which in turn dephosphorylates and activates NFAT (Rao et al., 1997). Truncation of the N terminus of Vav1 is accompanied by a decrease in PLCG1 phosphorylation and this inhibits calcium mobilization (Knyazhitsky et al., 2012). Role in cytoskeleton: PLCG1 plays a role in actin reorganization (Pei et al., 1996; Wells, 2000; Wang et al., 2007; Li et al., 2009). Role in adhesion and migration: PLCG1 mediates cell adhesion and migration through an undefined mechanism (Wang et al., 2007; Crooke et al., 2009). PLCG1 plays a role in integrin-mediated cell motility processes (Jones et al., 2005). Role in apoptosis: PLCG1 is proteolytically cleaved by group II caspases especially by caspase-3 and caspase-7 during apoptosis. This results in the loss of receptor-mediated tyrosine phosphorylation (Bae et al., 2000). PLCG1 plays a protective role in H2O2-induced PC12 cells death (Yuan et al., 2009). The Fas-mediated apoptosis requires endoplasmic reticulum-mediated calcium release in a mechanism Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) dependent on PLCG1 activation (Wozniak et al., 2006). Role in transformation: PLCG1 interacts with Middle tumor antigen (MT). The tyrosine phosphorylation level of PLCG1 is elevated in cells expressing wild type MT but not in cells expressing Tyr322→Phe MT (Su et al., 1995). Role in autoimmune symptoms: PLCG1 deficiency impairs the development and function regulatory cells (FoxP3+), causing inflammatory/autoimmune symptoms (Fu et al., 2010). Homology The protein contains eight domains, four of which are unique to PLCG family (Suh et al., 2008). The PLCG 'specific array' of domains, comprising a "split" PH domain flanking two tandem SH2 domains and one SH3 domain, is inserted between the two halves (X and Y) of the TIM-barrel catalytic domain (Bunney and Katan, 2011). Several other domains including two PH domains, one C2 domain and one EF hand motifs (Suh et al., 2008). Mutations Somatic 99 mutations have been described in the PLCG1 gene, according to the Catalogue of Somatic Mutations in Cancer (COSMIC) database. De novo mutation has been described in patients with Cutaneous T-cell lymphoma (CTCL): S345F (10/53 analyzed CTCL samples, 19%) (Vaqué et al., 2014). Implicated in Breast cancer Oncogenesis Overexpression of PLCG1 is a marker of development of metastases in breast cancer (Lattanzio et al., 2013). Loss of PLCG1 in part mimicked the effect of miR200b/miR-c/miR-429 overexpression in viability, apoptosis and EGF-driven cell invasion of breast cancer cells (Uhlmann et al., 2010). 669 PLCG1 (Phospholipase C, Gamma 1) Manso R signaling towards NFAT activation (Vaqué et al., 2014). Colorectal cancer Oncogenesis PLCG1 has a potencial role in colon cancer (Nomoto et al., 1995; Li et al., 2005; Reid et al., 2009). The activity of PLCG1 is reduced in STAT3 Y705F mutant colorectal cancer cells (Zhang et al., 2011), it shows that there is crosstalk between STAT3 and PLCG1 signaling pathways. Brain disorders Note Jang et al., 2013. Oncogenesis PLCG1 is highly expressed in brain. Abnormal expression and activation of PLCG1 appears in epilepsy (He et al., 2010), bipolar disorder (Løvlie et al., 2001), depression (Dwivedi et al., 2005), Huntington's disease (Giralt et al., 2009) and Alzheimer's disease (Shimohama et al., 1995). Prostate carcinoma Oncogenesis PLCG1 has a role in the regulation of PC3LN3 (human prostate carcinoma cells) cell adhesion that appears to be independent of its effects on tumour cell chemotactic migration and spreading in response to extracellular matrix (Peak et al., 2008). Myocardial dysfunction in sepsis Oncogenesis PLCG1 signaling induces cardiac TNF-alpha expression and myocardial dysfunction during Lipopolysaccharide (LPS) stimulation. Inhibition of PLCG1 decreased cardiac TNF-alpha expression and LPS-induced myocardial dysfunction was also attenuated (Peng et al., 2008). Gastric cancer Oncogenesis PLCG1 plays a role in RhoGDI2-mediated cisplatin resistance and cell invasion in gastric cancer (Cho et al., 2011). Squamous cell carcinoma (SCC) To be noted Oncogenesis PLCG1 is a downstream target of EGFR signaling. PLCG1 is required for EGFR-induced SCC cell mitogenesis (Xie et al., 2010). Note miR that target PLCG1: PLCG1 is target of different microRNAs, according to the bioinformatic algorithms microRNA (microRNA.org). Oral potentially malignant lesions (OPLs) References Oncogenesis PLCG1 is highly expressed in oral cancer lesions compared with normal oral mucosa (Ma et al., 2013). Nishibe S, Wahl MI, Wedegaertner PB, Kim JW, Rhee SG, Carpenter G. Selectivity of phospholipase C phosphorylation by the epidermal growth factor receptor, the insulin receptor, and their cytoplasmic domains. Proc Natl Acad Sci U S A. 1990 Jan;87(1):424-8 Esophageal adenocarcinoma Oncogenesis PLCG1-PKC-ERK pathway promotes proliferation and it is activated by VEGFR-2 in neoplastic Barrett's cells (Zhang et al., 2013). Smith RJ, Sam LM, Justen JM, Bundy GL, Bala GA, Bleasdale JE. 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Invest Ophthalmol Vis Sci. 2010 Dec;51(12):6803-9 Uhlmann S, Zhang JD, Schwäger A, Mannsperger H, Riazalhosseini Y, Burmester S, Ward A, Korf U, Wiemann S, Sahin O. miR-200bc/429 cluster targets PLCgamma1 and differentially regulates proliferation and EGF-driven invasion than miR-200a/141 in breast cancer. Oncogene. 2010 Jul 29;29(30):4297-306 Zhang Q, Yu C, Peng S, Xu H, Wright E, Zhang X, Huo X, Cheng E, Pham TH, Asanuma K, Hatanpaa KJ, Rezai D, Wang DH, Sarode V, Melton S, Genta RM, Spechler SJ, Souza RF. Autocrine VEGF signaling promotes proliferation of neoplastic Barrett's epithelial cells through a PLC-dependent pathway. Gastroenterology. 2014 Feb;146(2):461-72.e6 Xie Z, Chen Y, Liao EY, Jiang Y, Liu FY, Pennypacker SD. Phospholipase C-gamma1 is required for the epidermal growth factor receptor-induced squamous cell carcinoma cell mitogenesis. Biochem Biophys Res Commun. 2010 Jun 25;397(2):296-300 This article should be referenced as such: Bunney TD, Katan M. PLC regulation: emerging pictures for molecular mechanisms. Trends Biochem Sci. 2011 Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Manso R. PLCG1 (Phospholipase C, Gamma 1). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):668-672. 672 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Gene Section Review SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5) Cesare Indiveri, Lorena Pochini, Michele Galluccio, Mariafrancesca Scalise Department DiBEST (Biologia, Ecologia, Scienze della Terra) Unit of Biochemistry and Molecular Biotechnology, University of Calabria, 87036 Arcavacata di Rende, Italy (CI, LP, MG, MS) Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/SLC1A5ID42313ch19q13.html DOI: 10.4267/2042/54036 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology constituted by 1737 nucleotides and differs in the 5' UTR from the variant NM_005628. In NM_001145144 the translation starts downstream the third exon generating a shorter peptide of 313 aa. The third isoform NM_ 001145145 has 1927 nucleotides and lacks the first exon. It presents a different translation start at 5', coding a peptide of 339 amino acids. A longer transcript, XM_005259167, is reported only in NCBI database. It has been identified by automated computational analysis. More than 400 SNP(s), both in coding and non-coding regions of the SLC1A5 gene, are reported in dbSNP database (dbSNP). More than 40 are responsible of amino acid substitutions with unknown significance. Only the variant SLC1A5P17A (rs3027956) is associated with breast cancer (Savas et al., 2006). A region constituted by 907 bp upstream of the ASCT2 gene possesses promoter activity (Bungard and McGivan, 2004). In this region the following putative elements have been identified: an amino acid-regulatory element, a consensus site for binding of the transcription factor activator protein 1 (AP1) and a consensus binding sites for nuclear and hepatocyte nuclear factors. Abstract Review on human SLC1A5, with data on DNA/RNA, on the protein encoded and pathological and physiological implications. Identity Other names: AAAT, ASCT2, ATBO, M7V1, M7VS1, R16, RDRC HGNC (Hugo): SLC1A5 Location: 19q13.32 Local order: Orientation: minus strand. DNA/RNA Description The SLC1A5 gene, located at 19q13.3, counts 28692 nucleotides with 8 exons. It has been found in 56 different organisms (NCBI). The gene encodes a protein involved in sodium-dependent neutral amino acid transport (Kekuda et al., 1996; Pingitore et al., 2013). Transcription Three isoforms (transcripts) are reported either on NCBI and Ensembl databases for SLC1A5 human gene, deriving from different translation start. They differ in length, particularly at 5' extremity. The first variant NM_005628 represents the longest transcript, constituted by 2873 nucleotides and 8 exons. This transcript encodes a peptide of 541 amino acids. The second variant NM_001145144 is Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Pseudogene The gene is virtually present in all vertebrates. The better known orthologous of the human gene are those from rat, mouse and rabbit. Identity between the human and rat, mouse, rabbit are 79%, 82% and 85%, respectively. 673 SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5) Indiveri C, et al. Figure 1. Isoforms of SLC1A5 gene. The three isoforms are present in the minus strand of the chromosome 19 in position 19q13.3. NM_005628: isoform one, encodes for the longest peptide and is constituted by 8 exons; NM_001145144: isoform two, due to alternative splicing is characterized by only four exons; NM_ 001145145: isoform three presents seven exons. The nucleotide sequence is depicted as black lines. Coding nucleotides and untranslated (UTR) regions are indicated by red and white boxes, respectively. Exons are indicated by roman numbers. Protein Function Description Transport mediated by the human ASCT2 has been originally studied in intact cell systems overexpressing the transport protein (Kekuda et al., 1996; Kekuda et al., 1997). Recently, hASCT2 was over-expressed in the yeast P. pastoris, purified and reconstituted in artificial phospholipid vesicles (proteoliposomes), in absence of other interfering transporters. All experimental systems concur in demonstrating that hASCT2 is an obligate exchanger of neutral amino acid. This antiport requires the presence of extracellular Na+ which cannot be substituted by Li+ or K+. The Na+ ex:amino acidex stoichiometry of the human transporter is likely to be 1:1. Competition studies on 3H-glutamine, 3H-threonine or 3H-alanine transport performed in cells indicated that other potential substrates of hASCT2 are valine, leucine, serine, cysteine, asparagine, methionine, isoleucine, tryptophan, histidine, phenylalanine. While glutamate, lysine, arginine along with MeAIB [α(methylamino)isobutyric acid] and BCH [2aminobicyclo-(2,2,1)-heptane-2-carboxylic acid] are neither transported nor inhibit hASCT2. Experiments with radioactive compounds confirmed the competition data (Torres-Zamorano et al., 1998). In proteoliposomes, inhibition has been confirmed for most but not for all of the amino acids. Moreover, proteoliposome studies highlighted an asymmetric specificity for amino acids allowing to distinguish the amino acids inwardly transported (alanine, cysteine, valine, methionine) from those bi-directionally transported (glutamine, serine, asparagine, and threonine). 541 amino acids; molecular mass 56598,34 Da. Human SLC1A5 is a permease (membrane transporter). The 3D structure is not available. Homology modeling highlights a structure similar to that of the glutamate transporter of P. horikoshii (1XFH). Nand C-terminal ends are intracellular. Potential site of N-glycosylation and phosphorilation are predicted. In the structural model, at least one glycosylation site is extracellular and the phosphorilation sites are intracellular (Fig. 2). Expression Human SLC1A5 has been originally named ASCT2 from AlaSerCysTransporter2 or ATB0. The acronym ASCT2 is the most frequently used to designate this transport system. It is expressed in many tissues, including brain, (Bröer and Brookes, 2001; Deitmer et al., 2003; Gliddon et al., 2009). There is functional evidence of the expression of ASCT2 in kidney and intestine (Bode, 2001). Besides Caco-2 cells, apparently, also the HT-29 intestinal cell line functionally expresses ASCT2 (Kekuda et al., 1996; Kekuda et al., 1997). Poly(A)1 RNA isolated from several tissues of human origin revealed expression in placenta, lung, skeletal muscle, kidney, and pancreas (Kekuda et al., 1996). Localisation The protein is localized in the plasma membrane. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 674 SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5) Indiveri C, et al. Figure 2. Homology structural model of hASCT2. Ribbon diagram viewing of the transporter from the lateral side. The model was built using the glutamate transporter Glpth from Pyrococcus horikoshii crystal structure (1XFH) as the template by Modeller V9.13. The homology model was represented using SpdbViewer 4.01. Asn 163 and 212, predicted as glycosilation sites, are highlighted in blue; Ser 183, 261 and Thr 206, 207, 329, predicted as phosphorilation sites are highlighted in red and orange, respectively. Prediction according to Scan Prosite. The functional asymmetry was also confirmed by the kinetic analysis of [3H]glutamine/glutamine antiport: different Km values were measured on the external and internal sides of proteoliposomes, 0,097 and 1,8 mM, respectively. The SH reagents HgCl2, mersalyl and pOHMB potently inhibited hASCT2 mediated transport (Pingitore et al., 2013). The physiological role of hASCT2 consists in providing cells with some neutral amino acids exporting others on the basis of the metabolic need of cells consistently with the intra and extracellular amino acid concentrations. In brain, particularly, hASCT2 contributes to glutamine homeostasis of neurons and astrocytes. On the basis of experiments performed with animal models, it was hypothesized that hASCT2 mediates efflux of glutamine from astrocytes, a process that is critical for the functioning of the glutamate-glutamine cycle to recover synaptically released glutamate in exchange with glutamine efflux (Bröer et al., 1999). The glutamine-glutamate cycle has been shown also in placenta. Glutamine crosses the placenta and enters the fetal liver where it is deamidated to glutamate. About 90% of glutamate generated by the liver is taken up by the placenta and used in the metabolism. The glutamine-glutamate cycle between the placenta and the fetal liver is obligatory for the generation of NADPH in the placenta (Torres-Zamorano et al., 1998). Among other functions reported for hASCT2 there is the Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) regulation of mTOR pathway, translation and autophagy. The transporter regulates an increase in the intracellular concentration of glutamine which is then used by another plasma membrane transporter, named LAT1 (SLC7A5) (Galluccio et al., 2013) as efflux substrate to regulate the uptake of extracellular leucine with subsequent activation of mTORC1 (Nicklin et al., 2009). Moreover, it has been proposed that a group of retroviruses specifically uses the hASCT2 as a common cell surface receptor following a co-evolution phenomenon. The orthologous murine transporter mASCT2 is inactive as a viral receptor (Marin et al., 2003). Implicated in Molecular basis of cancerogenesis Note Tumor cells acquire altered metabolism. Due to these changes, the expression of membrane transporters involved in providing nutrients is altered. The plasma membrane transporter for glutamine ASCT2 has been clearly associated to cancer development and progression, together with another amino acid membrane transporter, LAT1 specific for glutamine and other neutral amino acids (Fuchs and Bode, 2005). The energetic needs of cancer cells are different from normal ones due to the Warburg effect. According to this phenomenon ATP derives from anaerobic glycolisis bypassing 675 SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5) mitochondrial function (Ganapathy et al., 2009). In this scenario glutamine provided by means of ASCT2 and LAT1 transport function sustains tumor growth and signaling through mTOR pathway (Nicklin et al., 2009). The importance of ASCT2 in this network is revealed by induction of apoptosis when silencing its gene in human hepatoma cells (Fuchs et al., 2004). In the following paragraphs specific examples of human cancers are reported. Breast cancer Note In breast cancer ASCT2 has been found over expressed together with other proteins related to glutamine metabolism like glutamminase and glutamate dehydrogenase (Kim et al., 2012). The study revealed that this metabolism is essential for sustaining breast cancer development and that the protein levels are different according to different subtypes of cancer. The subtype HER2 showed the highest level of glutamine related proteins and that the basal-like breast cancers are more dependent on glutamine compared to luminallikeones. Prostate cancer Note Tissue microarray technology (TMA) has been used for studying ASCT2 in normal prostatic tissue, in benign prostatic hyperplasia and in prostate adenocarcinoma. In particular, a negative prognosis and a shorter time of recurrence for adenocarcinoma were associated to hASCT2 expression. Moreover, a more aggressive behavior of adenocarcinoma is described (Li et al., 2003). Other diseases Note Due to importance of glutamine in cell metabolism and the chromosomal localization of SLC1A5 gene, several association studies have been conducted to ascertain the involvement of hASCT2 in pathologies like cystinuria, cystic fibrosis, schizophrenia, Hartnup disorder and pre-eclampsia. However, no genetic associations have been revealed. Colorectal carcinoma Note The expression of ASCT2 in colorectal carcinoma is normally associated to a decrease of percentage in patient survival (Witte et al., 2002). To be noted Note Aknowledgements: This work was supported by funds from: Programma Operativo Nazionale [PON 01_00937] "Modelli sperimentali Biotecnologici integrati per lo sviluppo e la selezione di molecole di interesse per la salute dell'uomo", Ministero Istruzione Università e Ricerca (MIUR). Neuroblastoma and glioma Note Neuroblastoma are childhood tumors very often benign. In some cases, however, neuroblastoma became malignant. One of the biological marker of this second category is the increased uptake of glutamine and other neutral aminoacids via ASCT2 (Wasa et al., 2002). Human glioma C6 cells have been demonstrated to mediate uptake of glutamine via ASCT2 (Dolinska et al., 2003). References Kekuda R, Prasad PD, Fei YJ, Torres-Zamorano V, Sinha S, Yang-Feng TL, Leibach FH, Ganapathy V. Cloning of the sodium-dependent, broad-scope, neutral amino acid transporter Bo from a human placental choriocarcinoma cell line. J Biol Chem. 1996 Aug 2;271(31):18657-61 Hepatoma Note Hepatocell carcinoma (HCC) is the most common malignant tumor of liver and one of the main cause of death. A study reported that higher rate of glutamine uptake via ASCT2 is a common feature of six examined hepatoma cell line (Bode et al., 2002; Fuchs et al., 2004). Kekuda R, Torres-Zamorano V, Fei YJ, Prasad PD, Li HW, Mader LD, Leibach FH, Ganapathy V. Molecular and functional characterization of intestinal Na(+)-dependent neutral amino acid transporter B0. Am J Physiol. 1997 Jun;272(6 Pt 1):G1463-72 Torres-Zamorano V, Leibach FH, Ganapathy V. Sodiumdependent homo- and hetero-exchange of neutral amino acids mediated by the amino acid transporter ATB degree. Biochem Biophys Res Commun. 1998 Apr 28;245(3):824-9 Lung cancer Note ASCT2 has been found over expressed in lung cancer by proteomic approach and then confirmed at molecular level. Pharmacologic and genetic targeting of ASCT2 decreased cell growth and viability in lung cancer cells, an effect mediated in part by mTOR signaling (Hassanein et al., 2013). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Indiveri C, et al. Bröer A, Brookes N, Ganapathy V, Dimmer KS, Wagner CA, Lang F, Bröer S. The astroglial ASCT2 amino acid transporter as a mediator of glutamine efflux. J Neurochem. 1999 Nov;73(5):2184-94 Bröer A, Wagner C, Lang F, Bröer S. Neutral amino acid transporter ASCT2 displays substrate-induced Na+ exchange and a substrate-gated anion conductance. Biochem J. 2000 Mar 15;346 Pt 3:705-10 676 SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5) Indiveri C, et al. Bode BP. Recent molecular advances in mammalian glutamine transport. J Nutr. 2001 Sep;131(9 Suppl):2475S85S; discussion 2486S-7S Fuchs BC, Bode BP. Amino acid transporters ASCT2 and LAT1 in cancer: partners in crime? Semin Cancer Biol. 2005 Aug;15(4):254-66 Bröer S, Brookes N. Transfer of glutamine between astrocytes and neurons. J Neurochem. 2001 May;77(3):705-19 Savas S, Schmidt S, Jarjanazi H, Ozcelik H. Functional nsSNPs from carcinogenesis-related genes expressed in breast tissue: potential breast cancer risk alleles and their distribution across human populations. Hum Genomics. 2006 Mar;2(5):287-96 Bode BP, Fuchs BC, Hurley BP, Conroy JL, Suetterlin JE, Tanabe KK, Rhoads DB, Abcouwer SF, Souba WW. Molecular and functional analysis of glutamine uptake in human hepatoma and liver-derived cells. Am J Physiol Gastrointest Liver Physiol. 2002 Nov;283(5):G1062-73 Ganapathy V, Thangaraju M, Prasad PD. Nutrient transporters in cancer: relevance to Warburg hypothesis and beyond. Pharmacol Ther. 2009 Jan;121(1):29-40 Wasa M, Wang HS, Okada A. Characterization of Lglutamine transport by a human neuroblastoma cell line. Am J Physiol Cell Physiol. 2002 Jun;282(6):C1246-53 Gliddon CM, Shao Z, LeMaistre JL, Anderson CM. Cellular distribution of the neutral amino acid transporter subtype ASCT2 in mouse brain. J Neurochem. 2009 Jan;108(2):372-83 Witte D, Ali N, Carlson N, Younes M. Overexpression of the neutral amino acid transporter ASCT2 in human colorectal adenocarcinoma. Anticancer Res. 2002 SepOct;22(5):2555-7 Nicklin P, Bergman P, Zhang B, Triantafellow E, Wang H, Nyfeler B, Yang H, Hild M, Kung C, Wilson C, Myer VE, MacKeigan JP, Porter JA, Wang YK, Cantley LC, Finan PM, Murphy LO. Bidirectional transport of amino acids regulates mTOR and autophagy. Cell. 2009 Feb 6;136(3):521-34 Deitmer JW, Bröer A, Bröer S. Glutamine efflux from astrocytes is mediated by multiple pathways. J Neurochem. 2003 Oct;87(1):127-35 Galluccio M, Pingitore P, Scalise M, Indiveri C. Cloning, large scale over-expression in E. coli and purification of the components of the human LAT 1 (SLC7A5) amino acid transporter. Protein J. 2013 Aug;32(6):442-8 Dolińska M, Dybel A, Zabłocka B, Albrecht J. Glutamine transport in C6 glioma cells shows ASCT2 system characteristics. Neurochem Int. 2003 Sep-Oct;43(4-5):5017 Li R, Younes M, Frolov A, Wheeler TM, Scardino P, Ohori M, Ayala G. Expression of neutral amino acid transporter ASCT2 in human prostate. Anticancer Res. 2003 JulAug;23(4):3413-8 Hassanein M, Hoeksema MD, Shiota M, Qian J, Harris BK, Chen H, Clark JE, Alborn WE, Eisenberg R, Massion PP. SLC1A5 mediates glutamine transport required for lung cancer cell growth and survival. Clin Cancer Res. 2013 Feb 1;19(3):560-70 Marin M, Lavillette D, Kelly SM, Kabat D. N-linked glycosylation and sequence changes in a critical negative control region of the ASCT1 and ASCT2 neutral amino acid transporters determine their retroviral receptor functions. J Virol. 2003 Mar;77(5):2936-45 Kim S, Kim do H, Jung WH, Koo JS. Expression of glutamine metabolism-related proteins according to molecular subtype of breast cancer. Endocr Relat Cancer. 2013 Jun;20(3):339-48 Bungard CI, McGivan JD. Glutamine availability upregulates expression of the amino acid transporter protein ASCT2 in HepG2 cells and stimulates the ASCT2 promoter. Biochem J. 2004 Aug 15;382(Pt 1):27-32 Pingitore P, Pochini L, Scalise M, Galluccio M, Hedfalk K, Indiveri C. Large scale production of the active human ASCT2 (SLC1A5) transporter in Pichia pastoris--functional and kinetic asymmetry revealed in proteoliposomes. Biochim Biophys Acta. 2013 Sep;1828(9):2238-46 Fuchs BC, Perez JC, Suetterlin JE, Chaudhry SB, Bode BP. Inducible antisense RNA targeting amino acid transporter ATB0/ASCT2 elicits apoptosis in human hepatoma cells. Am J Physiol Gastrointest Liver Physiol. 2004 Mar;286(3):G467-78 Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) This article should be referenced as such: Indiveri C, Pochini L, Galluccio M, Scalise M. SLC1A5 (solute carrier family 1 (neutral amino acid transporter), member 5). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):673-677. 677 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Gene Section Short Communication USB1 (U6 snRNA biogenesis 1) Elisa Adele Colombo Genetica Medica, Dipartimento di Scienze della Salute, Universita degli Studi di Milano, Italy (EAC) Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Genes/USB1ID44608ch16q21.html DOI: 10.4267/2042/54037 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology Abstract DNA/RNA C16orf57 alias USB1 is the gene which mutations underlie poikiloderma with neutropenia (PN) syndrome, a rare genodermatosis with autosomic recessive inheritance. PN patients have an increased risk to develop myelodysplasia and acute myeloid leukaemia in the second decade of life. In 2012, the protein encoded by USB1 has been recognised to be a 2H phosphodiesterase involved in the processing of U6 snRNA, but its action pathway and hence role in the pathogenesis of PN has not yet been elucidated. Description According to UCSC database (GRCh37/hg19, Feb.2009), USB1 gene maps in the region between 58035277 and 58055527 bp from pter of chromosome 16 with a centromeric-telomeric orientation. It spans 20 kb and is composed of seven exons (GI:305855061; NM_024598.3) (Fig.2). Transcription Two physiological isoforms, generated by alternative splicing (Fig. 2), have been detected in normal samples (leucocytes, keratinocytes, melanocytes and fibroblasts). The major transcript of 2282 nt (isoform 1, NM_024598.3) includes all the seven exons of the gene, while the shorter isoform of 1217 nt (NM_001204911.1) comprises the first three exons and an alternative terminal fourth exon located in IVS3 (Arnold et al., 2010). Identity Other names: C16orf57, EC 3.1.4., hUsb1, HVSL1, Mpn1, PN HGNC (Hugo): USB1 Location: 16q21 Figure 1. The region on chromosome 16q21 containing USB1 and its neighbouring genes ZNF139 (zinc finger protein 319) and MMP15 (matrix metalloproteinase 15) (UCSC database -GRCh37/hg19, Feb 2009). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 678 USB1 (U6 snRNA biogenesis 1) Colombo EA Figure 2. Schematic representation of exon-intron structure of USB1 and the two major transcripts resulting from alternative splicing of the two mutually exclusive exons 4. and serine residues (H120, S122, and H208, S210) which are essential for its catalytic activity. Recognition of these motifs by computational analysis of the protein sequence has predicted USB1 belongs to the 2H phosphodiesterase superfamily present in bacteria, archea and eukaryotes (Colombo et al., 2012). The protein has a globular architecture with two juxtaposed lobes with a pseudo two-fold symmetry separated by a central groove, which exposes the two HLSL motifs of the active site (Fig.3). Several additional transcripts, a few detected in cancer samples, are reported in the Ensembl database. Pseudogene No pseudogene for USB1 is known. Protein Description The crystal structure of the human USB1 protein, translated by isoform 1 mRNA has been recently resolved (Hilcenko et al., 2013). The main USB1 protein comprises 265 aa, while translation of isoform 4 mRNA predicts a 186 amino acid protein with a different C-terminus. The USB1 protein is characterized by two tetrapeptide motifs (HLSL), containing histidine Expression USB1 is ubiquitously expressed in humans (Volpi et al., 2010). The high evolutionary conservation of the protein is consistent with the housekeeping function of the gene. Figure 3. Ribbon model of the USB1 protein showing its globular symmetrical conformation with two lobes separated by a central groove that exposes the catalytic site containing the two HLSL motifs (encircled). The terminal lobe comprises both the Nand the C-termini. Both the terminal and transit lobe consist of antiparallel β-sheets and α-helices (modified from Colombo et al., 2012). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 679 USB1 (U6 snRNA biogenesis 1) Colombo EA Localisation Mutations A nuclear localization of USB1 has been demonstrated in HeLa cells (Mroczek et al., 2012); both nuclear and mitochondrial localizations have been observed for the yeast orthologue (Glatigny et al., 2011). Germinal Biallelic mutations in USB1 gene (OMIM*613276) cause poikiloderma with neutropenia syndrome (OMIM#604173). To date, 19 different "loss-of-function" mutations have been identified in 38 molecularly tested PN patients: 7 non-sense mutations, 6 out-of-frame deletions and 6 canonical splice site mutations. The latter also include the only missense mutation so far reported which however leads to exon skipping (Volpi et al., 2010). Recurrent mutations can be identified in patients of Navajo, Turkish and Caucasian origin attesting a founder effect (Colombo et al., 2012). Function Usb1 is a 3'-5' RNA exoribonuclease that trims the 3' end of the U6 snRNA leading to the formation of a terminal 2',3' cyclic phosphate. This posttranscriptionally modification influences U6 stability and recycling. Evidence has been obtained in yeast where Usb1 depletion leads to reduced levels of U6, generalized pre-mRNA splicing defects and shorter telomeres. In human use of PN cell lines confirmed that U6 is a substrate of USB1, but failed to reveal a splicing defect leaving unsolved how PN develops (Hilcenko et al., 2012; Mroczek et al., 2012; Shchepachev et al., 2012). Somatic No information is currently available on mutations of USB1 in sporadic cancers. Figure 4. Map across the USB1 gene of the currently known 19 mutations. Nonsense mutations are represented with a red hexagon, deletions with a yellow star and splicing mutations with a blue triangle. The Table lists for each mutation the intragenic position, the description (cDNA nomenclature) and the effect at the protein level. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 680 USB1 (U6 snRNA biogenesis 1) Colombo EA stratify the patients according to life-long cancer risk (myelodysplasia and solid tumours). Further studies focussing on the alternative transcript are necessary to establish the role of isoform 4 on PN pathogenesis and prognosis. Implicated in Poikiloderma with neutropenia syndrome (PN) Note The disease is caused by mutations affecting the gene represented in this entry. The clinical presentation of PN patients partially overlaps that of patients affected with RothmundThomson syndrome (RTS; OMIM#268400) and dyskeratosis congenita (DC; OMIM#613987, #613988, #613989, #615190, #224230). Disease Poikiloderma with neutropenia is a rare inherited genodermatosis characterized by skin alterations (poikiloderma, nail dystrophy, palmo-plantar hyperkeratosis), short stature and non cyclic neutropenia. In infancy, neutropenia is responsible of the recurrent infections, mainly of the respiratory system, observed in PN patients and, later in life, may lead to myelodysplastic syndrome and acute myeloid leukaemia. Squamous cell carcinoma has also been reported in PN patients. To date, 38 out of 66 PN patients described in literature have been molecularly tested and found to carry biallelic mutations of the USB1 gene. Most of the reported patients carry homozygous mutations, attesting inheritance by descent of the same ancestral mutation. Prognosis The knowledge of USB1 3D structure with the essential amino acid motifs of the catalytic site might enhance the prediction of USB1 mutation effects. All the mutations reported so far in PN patients (no. 19) interfere with USB1 function: 16 disrupt the catalytic activity due to the loss of one or both HLSL motifs, while the remaining 3 mutations, although not affecting the catalytically active tetrapeptide motifs destroy the internal symmetry of the protein. Owing to the restricted number of molecularly characterised PN patients no mutationphenotype correlations have emerged suitable to Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) References Arnold AW, Itin PH, Pigors M, Kohlhase J, BrucknerTuderman L, Has C. Poikiloderma with neutropenia: a novel C16orf57 mutation and clinical diagnostic criteria. Br J Dermatol. 2010 Oct;163(4):866-9 Volpi L, Roversi G, Colombo EA, Leijsten N, Concolino D, Calabria A, Mencarelli MA, Fimiani M, Macciardi F, Pfundt R, Schoenmakers EF, Larizza L. Targeted next-generation sequencing appoints c16orf57 as clericuzio-type poikiloderma with neutropenia gene. Am J Hum Genet. 2010 Jan;86(1):72-6 Glatigny A, Mathieu L, Herbert CJ, Dujardin G, Meunier B, Mucchielli-Giorgi MH. An in silico approach combined with in vivo experiments enables the identification of a new protein whose overexpression can compensate for specific respiratory defects in Saccharomyces cerevisiae. BMC Syst Biol. 2011 Oct 25;5:173 Colombo EA, Bazan JF, Negri G, Gervasini C, Elcioglu NH, Yucelten D, Altunay I, Cetincelik U, Teti A, Del Fattore A, Luciani M, Sullivan SK, Yan AC, Volpi L, Larizza L. Novel C16orf57 mutations in patients with Poikiloderma with Neutropenia: bioinformatic analysis of the protein and predicted effects of all reported mutations. Orphanet J Rare Dis. 2012 Jan 23;7:7 Mroczek S, Krwawicz J, Kutner J, Lazniewski M, Kuciński I, Ginalski K, Dziembowski A. C16orf57, a gene mutated in poikiloderma with neutropenia, encodes a putative phosphodiesterase responsible for the U6 snRNA 3' end modification. Genes Dev. 2012 Sep 1;26(17):1911-25 Shchepachev V, Wischnewski H, Missiaglia E, Soneson C, Azzalin CM. Mpn1, mutated in poikiloderma with neutropenia protein 1, is a conserved 3'-to-5' RNA exonuclease processing U6 small nuclear RNA. Cell Rep. 2012 Oct 25;2(4):855-65 Hilcenko C, Simpson PJ, Finch AJ, Bowler FR, Churcher MJ, Jin L, Packman LC, Shlien A, Campbell P, Kirwan M, Dokal I, Warren AJ. Aberrant 3' oligoadenylation of spliceosomal U6 small nuclear RNA in poikiloderma with neutropenia. Blood. 2013 Feb 7;121(6):1028-38 This article should be referenced as such: Colombo EA. USB1 (U6 snRNA biogenesis 1). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):678-681. 681 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Leukaemia Section Short Communication t(9;15)(p13;q24) PAX5/PML Jean-Loup Huret Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers, France (JLH) Published in Atlas Database: January 2014 Online updated version : http://AtlasGeneticsOncology.org/Anomalies/t0915p13q24ID1561.html DOI: 10.4267/2042/54038 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology Abstract Genes involved and proteins Short Communication on t(9;15)(p13;q24) PAX5/PML, with data on clinics, and the genes implicated. PAX5 Location 9p13.2 Protein 391 amino acids; from N-term to C-term, PAX5 contains: a paired domain (aa: 16-142); an octapeptide (aa: 179-186); a partial homeodomain (aa: 228-254); a transactivation domain (aa: 304359); and an inhibitory domain (aa: 359-391). Lineage-specific transcription factor; recognizes the concensus recognition sequence GNCCANTGAAGCGTGAC, where N is any nucleotide. Involved in B-cell differentiation. Entry of common lymphoid progenitors into the B cell lineage depends on E2A, EBF1, and PAX5; activates B-cell specific genes and repress genes involved in other lineage commitments. Activates the surface cell receptor CD19 and repress FLT3. Pax5 physically interacts with the RAG1/RAG2 complex, and removes the inhibitory signal of the lysine-9-methylated histone H3, and induces V-toDJ rearrangements. Genes repressed by PAX5 expression in early B cells are restored in their function in mature B cells and plasma cells, and PAX5 repressed (Fuxa et al., 2004; Johnson et al., 2004; Zhang et al., 2006; Cobaleda et al., 2007; Medvedovic et al., 2011). Identity Note The translocation is noted with various breakpoints on chromosome 15, ranging from q22 to q25 (this is reminiscent of the t(15;17) PML/RARA). Clinics and pathology Disease B-cell acute lymphoblastic leukemia (B-ALL) Epidemiology Two cases to date, a 9-month old girl and a 1.5-year old boy, both with a CD10+ ALL (Nebral et al., 2007; Nebral et al., 2009). Prognosis A patient remains in complete remission 84 months from diagnosis, while the other one had a testicular relapse 2 years after diagnosis and died. Cytogenetics Cytogenetics morphological The translocation was the sole abnormality. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 682 t(9;15)(p13;q24) PAX5/PML Huret JL PAX5/PML fusion protein. PML Fusion protein Location 15q24.1 Protein 882 amino-acids (aa) and shorter isoforms with distinct C terminus sequences; from N-term to Cterm, PML contains: a proline rich domain (aa 346); a zinc finger (RING finger type) (aa 57-92); two zinc fingers (B-box types) (aa 124-166 and aa 183-236); a coiled coil made of hydrophobic aa heptad repeats (aa 228-253); an interaction domain with PER2 (aa 448-555); a nuclear localization signal (aa 476-490); a proline rich domain (aa 504583); a serine rich domain (aa 506-540); and a sumo interaction motif (aa 556-562). The RING finger, B-boxes, and coiled-coil region form a tripartite motif known as the TRIM or the RBCC motif, and is associated with E3 ubiquitin ligase activity. PML is the organizer of nuclear domains called nuclear bodies, which recruit a wide variety of proteins, most often sumoylated. PML is involved in DNA damage response, cell division control, chromosome instability, and is a clock regulator via regulation of PER2 expression. PML has proapoptotic functions, induces senescence, inhibits angiogenesis and cell migration (Grignani et al., 1996; Chen et al., 2012; de Thé et al., 2012). Description 1099 amino acids. The predicted fusion protein contains the DNA binding paired domain of PAX5 (260 aa from PAX5) and most of PML (839 aa). References Grignani F, Testa U, Rogaia D, Ferrucci PF, Samoggia P, Pinto A, Aldinucci D, Gelmetti V, Fagioli M, Alcalay M, Seeler J, Grignani F, Nicoletti I, Peschle C, Pelicci PG. Effects on differentiation by the promyelocytic leukemia PML/RARalpha protein depend on the fusion of the PML protein dimerization and RARalpha DNA binding domains. EMBO J. 1996 Sep 16;15(18):4949-58 Fuxa M, Skok J, Souabni A, Salvagiotto G, Roldan E, Busslinger M. Pax5 induces V-to-DJ rearrangements and locus contraction of the immunoglobulin heavy-chain gene. Genes Dev. 2004 Feb 15;18(4):411-22 Johnson K, Pflugh DL, Yu D, Hesslein DG, Lin KI, Bothwell AL, Thomas-Tikhonenko A, Schatz DG, Calame K. B cellspecific loss of histone 3 lysine 9 methylation in the V(H) locus depends on Pax5. Nat Immunol. 2004 Aug;5(8):85361 Zhang Z, Espinoza CR, Yu Z, Stephan R, He T, Williams GS, Burrows PD, Hagman J, Feeney AJ, Cooper MD. Transcription factor Pax5 (BSAP) transactivates the RAGmediated V(H)-to-DJ(H) rearrangement of immunoglobulin genes. Nat Immunol. 2006 Jun;7(6):616-24 Cobaleda C, Schebesta A, Delogu A, Busslinger M. Pax5: the guardian of B cell identity and function. Nat Immunol. 2007 May;8(5):463-70 Result of the chromosomal anomaly Nebral K, König M, Harder L, Siebert R, Haas OA, Strehl S. Identification of PML as novel PAX5 fusion partner in childhood acute lymphoblastic leukaemia. Br J Haematol. 2007 Oct;139(2):269-74 Hybrid gene Nebral K, Denk D, Attarbaschi A, König M, Mann G, Haas OA, Strehl S. Incidence and diversity of PAX5 fusion genes in childhood acute lymphoblastic leukemia. Leukemia. 2009 Jan;23(1):134-43 Description Fusion of PAX5 exon 6 to PML exon 2. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 683 t(9;15)(p13;q24) PAX5/PML Huret JL Medvedovic J, Ebert A, Tagoh H, Busslinger M. Pax5: a master regulator of B cell development and leukemogenesis. Adv Immunol. 2011;111:179-206 de Thé H, Le Bras M, Lallemand-Breitenbach V. The cell biology of disease: Acute promyelocytic leukemia, arsenic, and PML bodies. J Cell Biol. 2012 Jul 9;198(1):11-21 Chen RH, Lee YR, Yuan WC. The role of PML ubiquitination in human malignancies. J Biomed Sci. 2012 Aug 30;19:81 This article should be referenced as such: Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Huret JL. t(9;15)(p13;q24) PAX5/PML. Atlas Cytogenet Oncol Haematol. 2014; 18(9):682-684. 684 Genet Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Leukaemia Section Short Communication t(1;9)(p13;p12) PAX5/HIPK1 Jean-Loup Huret Genetics, Dept Medical Information, University of Poitiers, CHU Poitiers Hospital, F-86021 Poitiers, France (JLH) Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Anomalies/t0109p13p12ID1557.html DOI: 10.4267/2042/54039 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology degradation of proteins) (aa 892-972), SUMO interaction motifs, required for nuclear localization and kinase activity (aa 902-926), a domain interacting with DAB2IP/MAP3K5 (aa 973-1209), a histidine-rich region (aa 1086-1154), and a tyrosine-rich region (aa 1175-1209) (YH region). The lysine residues at the sumoylation motifs are the following: K25, K317, K440, K556, and K1202 (Kim et al., 1998; Li et al., 2005; Swiss-Prot). First identified as a nuclear serine/threonine kinase. Homeodomain-interacting protein kinase. HIPK1 positively or negatively modulate signaling pathways controlling cell proliferation and/or apoptosis (review in Rinaldo et al., 2008). HIPK1 and HIPK2 act cooperatively as corepressors in the transcriptional activation of angiogenic genes, including MMP10 (11q22.2) and VEGFA (6p21.1), that are critical for the early stage of vascular development (Shang et al., 2013). HIPK1 regulates the p53 signaling pathway. PARK7 (also called DJ-1, 1p36.23), a protein also linked to the p53 signaling pathway, is able to induce HIPK1 degradation. HIPK1 directly phophorylates TP53 on its serine15. Serine 15 phosphorylation induces a rise in CDKN1A (p21, 6p21.2) expression and cell cycle arrest (Rey et al., 2013). HIPK1 phosphorylates DAXX (6p21.32), a protein which interacts with PML (15q24.1), the organizer of nuclear bodies (Ecsedy et al., 2003), and which relocalizes from the nucleus to the cytoplasm in response to stress. During glucose deprivation, a pathway involving MAP3K5 (also called ASK1, 6q23.3), -> MAP2K4 (SEK1, 17p12) -> MAPK8 (JNK1, 10q11.22) -> HIPK1 is activated, and DAXX is relocated in the cytoplasm (Song and Lee, 2003). Abstract Short communication on t(1;9)(p13;p12) PAX5/HIPK1, with data on clinics, and the genes implicated. Clinics and pathology Disease B-cell acute lymphoblastic leukemia (B-ALL) Epidemiology Only one case to date, a 3-year old boy with a CD10+ ALL (Nebral et al., 2009). Prognosis The patient was noted at an intermediate risk, and was in complete remission 6 months after diagnosis. Genes involved and proteins HIPK1 Location 1p13.2 Protein 1210 amino acids (aa). From N-term to C-term, contains a protein kinase domain (aa 190-518), a nucleotide binding motif (aa 196-204), a domain interacting with DAB2IP (AIP1, or AF9q34, 9q33.2) (aa 518-889), a nuclear localization signal (aa 844-847), a region interacting with TP53 (aa 885-1093), a nuclear speckle retention signal (aa 887-992), (corresponding to aa 860-967 in HIPK2), a PEST domain (enriched in proline (P), glutamate (E), serine (S), and threonine (T), expedite the Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 685 t(1;9)(p13;p12) PAX5/HIPK1 Huret JL t(1;9)(p13;p13) PAX5/HIPK1 fusion protein. TNF (TNF-alpha, 6p21.33) induces desumoylation and cytoplasm translocation of HIPK1 leading to apoptosis (Li et al., 2005). HIPK1 and HIPK2 bind HOX genes homeodomains and regulate their expression, as well as PAX1 (20p11.22) and PAX3 (2q36.1) transcription (Isono et al., 2006). HIPK1 and HIPK2 phosphorylates EP300 (22q13.2) and RUNX1 (21q22.12) during embryonic development, and Hipk1/Hipk2-deficient mice show defective definitive hematopoiesis (Aikawa et al., 2006). HIPK1 phosphorylates MYB (6q23.3), a transcriptional activator essential for the establishment of haemopoiesis, and causes repression of MYB activation (Matre et al., 2009). HIPK1 interacts with DVL1 (1p36.33) and TCF3 (E2A, 19p13.3) and regulates Wnt/b-catenin target genes during early embryonic development (Louie et al., 2009). HIPK1 is highly overexpressed in colorectal carcinomas compared with healthy mucosa. The highest peak of HIPK1 expression occurred at early stages and decreased in latter stages. HIPK1 appeared to be induced as a defense mechanism to fight against intern deregulations and stressful conditions, rather than produced by the cancer cells as an indispensable factor for tumor evolution (Rey et al., 2013). HIPK1 is expressed only in invasive breast cancer cells with three different subcellular localization, associated with different tumor histopathologic characteristics (Park et al., 2012). Protein 391 amino acids; from N-term to C-term, PAX5 contains: a paired domain (aa: 16-142); an octapeptide (aa: 179-186); a partial homeodomain (aa: 228-254); a transactivation domain (aa: 304359); and an inhibitory domain (aa: 359-391). Lineage-specific transcription factor; recognizes the concensus recognition sequence GNCCANTGAAGCGTGAC, where N is any nucleotide. Involved in B-cell differentiation. Entry of common lymphoid progenitors into the B cell lineage depends on E2A, EBF1, and PAX5; activates B-cell specific genes and repress genes involved in other lineage commitments. Activates the surface cell receptor CD19 and repress FLT3. Pax5 physically interacts with the RAG1/RAG2 complex, and removes the inhibitory signal of the lysine-9methylated histone H3, and induces V-to-DJ rearrangements. Genes repressed by PAX5 expression in early B cells are restored in their function in mature B cells and plasma cells, and PAX5 repressed (Fuxa et al., 2004; Johnson et al., 2004; Zhang et al., 2006; Cobaleda et al., 2007; Medvedovic et al., 2011). PAX5 Hybrid gene Location 9p13.2 Description Fusion of PAX5 exon 5 to HIPK1 exon 9. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Result of the chromosomal anomaly 686 t(1;9)(p13;p12) PAX5/HIPK1 Huret JL homeodomain-interacting protein kinases hipk1 and hipk2 in the mediation of cell growth in response to morphogenetic and genotoxic signals. Mol Cell Biol. 2006 Apr;26(7):2758-71 Fusion protein Description 751 amino acids. The predicted fusion protein contains the DNA binding paired domain of PAX5 and the nuclear localization signal, the region interacting with TP53, the nuclear speckle retention signal, the PEST domain, the SUMO interaction motifs, the domain interacting with DAB2IP/MAP3K5, and theYH region from HIPK1. Zhang Z, Espinoza CR, Yu Z, Stephan R, He T, Williams GS, Burrows PD, Hagman J, Feeney AJ, Cooper MD. Transcription factor Pax5 (BSAP) transactivates the RAGmediated V(H)-to-DJ(H) rearrangement of immunoglobulin genes. Nat Immunol. 2006 Jun;7(6):616-24 Cobaleda C, Schebesta A, Delogu A, Busslinger M. Pax5: the guardian of B cell identity and function. Nat Immunol. 2007 May;8(5):463-70 References Rinaldo C, Siepi F, Prodosmo A, Soddu S. HIPKs: Jack of all trades in basic nuclear activities. Biochim Biophys Acta. 2008 Nov;1783(11):2124-9 Kim YH, Choi CY, Lee SJ, Conti MA, Kim Y. Homeodomain-interacting protein kinases, a novel family of co-repressors for homeodomain transcription factors. J Biol Chem. 1998 Oct 2;273(40):25875-9 Louie SH, Yang XY, Conrad WH, Muster J, Angers S, Moon RT, Cheyette BN. Modulation of the beta-catenin signaling pathway by the dishevelled-associated protein Hipk1. PLoS One. 2009;4(2):e4310 Ecsedy JA, Michaelson JS, Leder P. Homeodomaininteracting protein kinase 1 modulates Daxx localization, phosphorylation, and transcriptional activity. Mol Cell Biol. 2003 Feb;23(3):950-60 Matre V, Nordgård O, Alm-Kristiansen AH, Ledsaak M, Gabrielsen OS. HIPK1 interacts with c-Myb and modulates its activity through phosphorylation. Biochem Biophys Res Commun. 2009 Oct 9;388(1):150-4 Song JJ, Lee YJ. Role of the ASK1-SEK1-JNK1-HIPK1 signal in Daxx trafficking and ASK1 oligomerization. J Biol Chem. 2003 Nov 21;278(47):47245-52 Nebral K, Denk D, Attarbaschi A, König M, Mann G, Haas OA, Strehl S. Incidence and diversity of PAX5 fusion genes in childhood acute lymphoblastic leukemia. Leukemia. 2009 Jan;23(1):134-43 Fuxa M, Skok J, Souabni A, Salvagiotto G, Roldan E, Busslinger M. Pax5 induces V-to-DJ rearrangements and locus contraction of the immunoglobulin heavy-chain gene. Genes Dev. 2004 Feb 15;18(4):411-22 Medvedovic J, Ebert A, Tagoh H, Busslinger M. Pax5: a master regulator of B cell development and leukemogenesis. Adv Immunol. 2011;111:179-206 Johnson K, Pflugh DL, Yu D, Hesslein DG, Lin KI, Bothwell AL, Thomas-Tikhonenko A, Schatz DG, Calame K. B cellspecific loss of histone 3 lysine 9 methylation in the V(H) locus depends on Pax5. Nat Immunol. 2004 Aug;5(8):85361 Park BW, Park S, Koo JS, Kim SI, Park JM, Cho JH, Park HS. Homeodomain-interacting protein kinase 1 (HIPK1) expression in breast cancer tissues. Jpn J Clin Oncol. 2012 Dec;42(12):1138-45 Li X, Zhang R, Luo D, Park SJ, Wang Q, Kim Y, Min W. Tumor necrosis factor alpha-induced desumoylation and cytoplasmic translocation of homeodomain-interacting protein kinase 1 are critical for apoptosis signal-regulating kinase 1-JNK/p38 activation. J Biol Chem. 2005 Apr 15;280(15):15061-70 Rey C, Soubeyran I, Mahouche I, Pedeboscq S, Bessede A, Ichas F, De Giorgi F, Lartigue L. HIPK1 drives p53 activation to limit colorectal cancer cell growth. Cell Cycle. 2013 Jun 15;12(12):1879-91 Shang Y, Doan CN, Arnold TD, Lee S, Tang AA, Reichardt LF, Huang EJ. Transcriptional corepressors HIPK1 and HIPK2 control angiogenesis via TGF-β-TAK1-dependent mechanism. PLoS Biol. 2013;11(4):e1001527 Aikawa Y, Nguyen LA, Isono K, Takakura N, Tagata Y, Schmitz ML, Koseki H, Kitabayashi I. Roles of HIPK1 and HIPK2 in AML1- and p300-dependent transcription, hematopoiesis and blood vessel formation. EMBO J. 2006 Sep 6;25(17):3955-65 This article should be referenced as such: Isono K, Nemoto K, Li Y, Takada Y, Suzuki R, Katsuki M, Nakagawara A, Koseki H. Overlapping roles for Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Huret JL. t(1;9)(p13;p12) PAX5/HIPK1. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):685-687. 687 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Solid Tumour Section Short Communication Lung: t(6;12)(q22;q14.1) LRIG3/ROS1 in lung adenocarcinoma Kana Sakamoto, Yuki Togashi, Kengo Takeuchi Pathology Project for Molecular Targets, The Cancer Institute, Japanese Foundation for Cancer Research, Tokyo, Japan (KS, YT, KT) Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Tumors/t0612q22q14inLungID6496.html DOI: 10.4267/2042/54040 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology UFT. Although not administered in this case, Crizotinib and other ALK inhibitors have been reported to be effective in lung cancers with ROS1 translocations (Bergethon et al., 2012; Shaw et al., 2012). Abstract Short communication on t(6;12)(q22;q14.1) LRIG3/ROS1 in lung adenocarcinoma with data on clinics. Prognosis Clinics and pathology With 5 years of follow-up, the patient was alive without relapse. Disease Lung adenocarcinoma Genes involved and proteins Epidemiology ROS1 translocations are found in 0.9 to 1.7% of non small cell lung carcinomas and the majority of the cases are adenocarcinoma (Bergethon et al., 2012; Davies et al., 2012; Takeuchi et al., 2012). Multiple fusion partners have been identified and LRIG3 is one of them. As for LRIG3-ROS1, only one case, a 57-year-old Japanese male patient, has been reported to date (Takeuchi et al., 2012). LRIG3 Location 12q14.1 DNA / RNA Leucine-Rich Repeats And Immunoglobulin-Like Domains Protein 3. Clinics ROS1 The patient had a 5 pack year of smoking history and was diagnosed as having stage 1A lung adenocarcinoma. Location 6q22 DNA / RNA C-Ros Oncogene 1, Receptor Tyrosine Kinase. Pathology This case showed moderately differentiated micropapillary pattern. A mucinous cribriform pattern which is frequently seen in cancers with kinase fusions was not found. This case was negative for EGFR and KRAS mutations as with the most cases harboring ROS1 gene fusions. Result of the chromosomal anomaly Hybrid Gene Treatment Transcript LRIG3-ROS1 fusion transcript was detected. The primary tumor was surgically removed and the patient received post-operative chemotherapy with Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 688 Lung: t(6;12)(q22;q14.1) LRIG3/ROS1 in lung adenocarcinomaSakamoto K, et al. A. The schematic structure of LRIG3, ROS1, and LRIG3-ROS1 proteins and the cDNA sequence around the fusion point: Exon 16 of LRIG3 fused to exon 35 of ROS1. The break point of ROS1 allows the resulting fusion protein to retain the kinase domain (red). LRIG3 contains a transmembrane domain (orange). B: RT-PCR confirmation of LRIG3-ROS1 fusion: Lane M and N represent the size standard (20-bp ladder) and the non-template control, respectively. C. Fusion FISH analysis: A fusion signal (yellow) was observed in consequence of the fusion of LRIG3 (red) and ROS1 (green). Detection A 218 bp cDNA fragment harboring the fusion point can be detected with LRIG3 forward primer (5'-ACACAGATGAGACCAACTTGC-3') and ROS1 reverse primer (5'CACTGTCACCCCTTCCTTG-3'). Oncogenesis The oncogenicity of LRIG3-ROS1 fusion was proven in a focus formation assay and a nude mouse tumorigenicity assay (Takeuchi et al., 2012). Fusion Protein Bergethon K, Shaw AT, Ou SH, Katayama R, Lovly CM, McDonald NT, Massion PP, Siwak-Tapp C, Gonzalez A, Fang R, Mark EJ, Batten JM, Chen H, Wilner KD, Kwak EL, Clark JW, Carbone DP, Ji H, Engelman JA, MinoKenudson M, Pao W, Iafrate AJ. ROS1 rearrangements define a unique molecular class of lung cancers. J Clin Oncol. 2012 Mar 10;30(8):863-70 References Description The fusion protein encompasses the constitutive activation of ROS1 tyrosine kinase. However, the mechanism of it is largely unknown. The role of LRIG3 here has not been clarified. LRIG3 protein does not contain a coiled-coil domain as in the case with most of the other ROS1 fusion partners (Takeuchi et al., 2012). In respect of the downstream signaling, several growth and survival signaling pathways which are common to other receptor tyrosine kinases have been shown to be involved. These include PI3K/AKT, JAK/STAT3, RAS/MAPK/ERK, VAV3, and SHP-1 and SHP-2 pathways (Chin et al., 2012; Davies and Doebele, 2013). Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Chin LP, Soo RA, Soong R, Ou SH. Targeting ROS1 with anaplastic lymphoma kinase inhibitors: a promising therapeutic strategy for a newly defined molecular subset of non-small-cell lung cancer. J Thorac Oncol. 2012 Nov;7(11):1625-30 Davies KD, Le AT, Theodoro MF, Skokan MC, Aisner DL, Berge EM, Terracciano LM, Cappuzzo F, Incarbone M, Roncalli M, Alloisio M, Santoro A, Camidge DR, VarellaGarcia M, Doebele RC. Identifying and targeting ROS1 gene fusions in non-small cell lung cancer. Clin Cancer Res. 2012 Sep 1;18(17):4570-9 Shaw AT, Camidge DR, Engelman JA, Solomon BJ, Kwak EL, Clark JW, Salgia R, Shapiro G, Bang YJ, Tan W, Tye 689 Lung: t(6;12)(q22;q14.1) LRIG3/ROS1 in lung adenocarcinomaSakamoto K, et al. L, Wilner KD, Stephenson P, Varella-Garcia M, Bergethon K, Iafrate AJ, Ou SHI.. Clinical activity of crizotinib in advanced non-small cell lung cancer (NSCLC) harboring ROS1 gene rearrangement. J Clin Oncol 2012;30:(suppl; abstr 7508). Davies KD, Doebele RC.. Molecular pathways: ROS1 fusion proteins in cancer. Clin Cancer Res. 2013 Aug 1;19(15):4040-5. doi: 10.1158/1078-0432.CCR-12-2851. Epub 2013 May 29. (REVIEW) This article should be referenced as such: Takeuchi K, Soda M, Togashi Y, Suzuki R, Sakata S, Hatano S, Asaka R, Hamanaka W, Ninomiya H et al.. RET, ROS1 and ALK fusions in lung cancer. Nat Med. 2012 Feb 12;18(3):378-81. doi: 10.1038/nm.2658. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Sakamoto K, Togashi Y, Takeuchi K. Lung: t(6;12)(q22;q14.1) LRIG3/ROS1 in lung adenocarcinoma. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):688690. 690 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Deep Insight Section The tumour suppressor function of the scaffolding protein spinophilin Denis Sarrouilhe, Véronique Ladeveze Laboratoire de Physiologie Humaine, Faculte de Medecine et Pharmacie, Universite de Poitiers, 6 rue de la Miletrie, Bat D1, TSA 51115, 86073 Poitiers, Cedex 9, France (DS), Laboratoire de Genetique Moleculaire de Maladies Rares, Universite de Poitiers, UFR SFA, Pole Biologie Sante, Bat B36, TSA 51106, 86073 Poitiers, Cedex 9, France (VL) Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Deep/SpinophilinID20133.html DOI: 10.4267/2042/54041 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology Abstract Spinophilin is a scaffolding protein with modular domains that govern its interaction with a large number of cellular proteins. The Spinophilin gene locus is localized at chromosome 17q21, a chromosomal region frequently affected by genomic instability in different human tumours. The scaffolding protein interacts with the tumour-suppressor ARF which has suggested a role for Spinophilin in cell growth. More recently, in vitro and in vivo studies demonstrated that Spinophilin is a new tumour suppressor acting via the regulation of pRb. A clear downregulation of Spinophilin is found in several human cancer types. Moreover, Spinophilin loss is associated with a poor patient prognosis in carcinoma. Currently, there are controversial findings regarding a functional relationship between Spinophilin and p53 in cell cycle regulation and in carcinogenesis. Here we present the available data regarding Spinophilin function as a tumour suppressor. Actin-Binding proteIN), and the latter was further identified as Spn (Nakanishi et al., 1997). Spn is expressed ubiquitously while neurabin 1 is expressed almost exclusively in neuronal cells. Spn exhibits the characteristics of scaffolding proteins with multiple protein interaction domains (Allen et al., 1997; Sarrouilhe et al., 2006). Scaffolding proteins link signalling enzymes, substrates and potential effectors (such as channels, receptors) into a multiprotein signalling complex that may be anchored to the cytoskeleton. In the years after this discovery, the spectrum of Spn partners and functions has expanded but has remained mostly in the field of neurobiology (Sarrouilhe et al., 2006). Spn has been implicated in the pathophysiology of several central nervous system (CNS) diseases, among which are Parkinson's disease, schizophrenia and mood disorders (Law et al., 2004; Brown et al., 2005). Spn is highly enriched at the synaptic membrane in dendritic spines, the site of excitatory neurotransmission and thus may control PP1 functions during synaptic activity (Ouimet et al., Key words CaSR, G protein-coupled receptor, signaling 1- Introduction Protein phosphatase 1 (PP1) is a widespread expressed phosphoSerine/phosphoThreonine PP involved in many cellular processes (Ceulemans and Bollen, 2004). There are four isoforms of PP1 catalytic subunit (PP1c): PP1α, PP1β, PP1γ1 and PP1γ2, the latter two arising through alternative splicing (Sasaki et al., 1990). PP1c can form complexes with up to 50 regulatory subunits converting the enzyme into many different forms, which have distinct substrates specificities, restricted subcellular locations and diverse regulations (Cohen, 2002). In late 1990s, a novel PP1c binding protein that is a potent modulator of PP1 activity was characterized in rat brain and named spinophilin (Spn) (Allen et al., 1997). In the same time, two novel actin filament-binding proteins were purified from rat brain and named neurabin 1 and neurabin 2 (NEURal tissue-specific- Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 691 The tumour suppressor function of the scaffolding protein spinophilin 2004). Spn regulates plasticity at the postsynaptic density (PSD) by targeting PP1c to α-amino-3hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and N-methyl-D-aspartic acid (NMDA) receptors, promoting their down regulation by dephosphorylation and thus regulating the efficiency of post-synaptic glutamatergic neurotransmission. Spn and neurabin1 play different roles in hippocampal and striatal synaptic plasticity. Spn is involved in long-term depression (LTD) but not in long-term potentiation (LTP) whereas neurabin 1 contributes selectively to LTP but not LTD (Feng et al., 2000; Allen et al., 2006; Wu et al., 2008). In the same way, the two scaffolding proteins form a functional pair of opposing regulators that reciprocally regulate signalling intensity by some seven-transmembrane domain receptors (Wang et al., 2007). Thus, an emerging notion is that Spn and neurabin 1 may differentially affect their target proteins and perform quite distinctive function in cell. Morphological studies have established that Spn is enriched at plasma membrane of cells although the protein is also expressed widely throughout the cytoplasm (Smith et al., 1999; Richman et al., 2001; Tsukada et al., 2003). Spn, which is expressed partly in the nucleus in mammalian cells, interacts in vitro and in vivo with the tumor-suppressor ARF (Alternative Reading Frame). Moreover, a role for Spn in cell growth was suggested, and this effect was enhanced by the interaction between Spn and ARF (Vivo et al., 2001). More recent studies showed that Spn is a new tumour suppressor and that a clear downregulation of this protein is found in several cancer types (Carnero, 2012). Furthermore, Spn loss is associated with poor patient prognosis in carcinomas (Sarrouilhe, 2014). This review aims to outline the state of knowledge regarding Spn function in carcinogenesis. domains, a PSD95/DLG/zo-1 (PDZ) and three coiled-coil domains. Figure 2 provides a schematic diagram of the main Spn structural domains. In the five species of the Figure 1, the coiled-coil region has high identity with only one variation detected in Cricetulus griseus. The PDZ domain, the pentapeptide motif of PP1c -binding domain and the sextapeptide allowing the binding selectivity of PP1c isoforms, present the same identity. Moreover, the phosphoSer are conserved except the Ser-177 which is only detected in rat. Being not detected in mouse (G as in primates), Ser-177 is not a consequence of the rodent-specific high substitution rate. Spn has been isolated from rat brain as a protein interacting with F-actin (Satoh et al., 1998). Its Factin-binding domain determined to be amino acids 1-154 is both necessary and sufficient to mediate actin polymers binding and cross-linking. Nuclear Magnetic Resonance (NMR) and circular dichroism (CD) spectroscopy studies showed that Spn F-actin-binding domain is intrinsically unstructured and that upon binding to F-actin it adopts a more ordered structure (a phenomenom also called folding-upon-binding). Another actin binding property, namely a F-actin pointed end capping activity was recently proposed for this domain (Schüler and Peti, 2007). Spn, PP1c and Factin can form a trimeric complex in vitro. A receptor-interacting domain, located between amino acids 151-444, interacts with the third intracellular loop (3i) of various seven transmembrane domain receptors (Smith et al., 1999; Richman et al., 2001) such as the dopamine D2 receptor (D2R), some subtypes of the αadrenergic (AR) and muscarinic-acetylcholine (mAchR) receptors. The primary PP1c-binding domain is located within residues 417-494 of Spn and this domain contains a pentapeptide motif (R-K-I-H-F) between amino acids 447 and 451 that is conserved in other PP1c regulatory subunits. A domain C-terminal to this canonical PP1-binding motif, located within amino acids 464 and 470, is essential for PP1 isoform selectivity in vitro and for selective targeting in cells (Carmody et al., 2008). Recently, the 3-dimentional structure of the PP1/Spn holoenzyme was determined. Spn is an unstructured protein in its unbound state that undergoes a folding transition upon interaction with PP1c into a single, stable conformation. The scaffolding protein binds to PP1c and blocks some potential substrate binding sites without altering its active site, then didacting substrate specificity of the enzyme (Ragusa et al., 2010). A further study showed that the PP1/Spn holoenzyme is dynamic in solution. 2- Spinophilin structure The primate (homo sapiens and Callithrix jacchus) Spn proteins contain 815 amino acids whereas the rodent Spn (rattus norvegicus and mus musculus) have 817amino acids. These sequences are very similar, with few amino acids substitutions compared to the human sequence in C-terminus but the N-terminus is more variable even if the variability is weak (Figure 1). Consequently, few differences are observed when we compared these sequences to the human one: the rat and human Spn proteins share 96% sequence identity (Allen et al., 1997; Vivo et al., 2001). In Cricetulus griseus, the sequence is shorter than the others: 631amino acids. Gene analysis and biochemical approaches have contributed to define in Spn a number of distinct modular domains. This 130 kDa protein contains one F-actin-, a receptor- and a PP1c- binding Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Sarrouilhe D, Ladeveze V 692 The tumour suppressor function of the scaffolding protein spinophilin Sarrouilhe D, Ladeveze V Figure 1. Alignment of amino acid sequences of spinophilin in different species. Blast and Align programs via UniProt site were used. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 693 The tumour suppressor function of the scaffolding protein spinophilin Sarrouilhe D, Ladeveze V Figure 2. Schematic drawing of spinophilin structure. The canonical protein phosphatase 1-binding domain is located within amino acids 447 and 451 in spinophilin. signalling protein (like RGS8), guanine nucleotide exchange factors (like kalirin 7), membrane receptors [like the α-ARs, m-AChRs, D2R, δ- and µ-opioid receptors (OR) and cholecystokinin (CCK) receptors], and other proteins like ions channels [The transient receptor potential canonical (TRPC), the type 2 ryanodine receptor (RYR2)], TGN38 and ARF. Shortly after the cloning of Spn as a novel PP1cbinding protein, another laboratory cloned this protein based on its ability to bind to F-actin (Satoh et al., 1998). Recombinant Spn and neurabin 1 interacted with each other when co-expressed in cells. On the other hand, recombinant Spn was shown to form homodimers, trimers or tetramers by interaction between coiled-coil domains. Spn homomeric complexes are thought to contribute to its actincross-linking activity (Satoh et al., 1998). Doublecortin (DCX) is a microtubule-associated protein that can induce microtubule polymerization and stabilize microtubules filaments. Immunoprecipitation experiments with brain extracts showed that Spn and DCX interact incultured cells (Tsukada et al., 2003). In vitro assays showed that DCX also binds to and bundles F-actin, suggesting that the protein crosslinks microtubules and F-actin. The distribution of DCX between the two cytoskeletons can be regulated by Spn and by phosphorylation of DCX and it was proposed that Spn could localize and enhance the binding of phosphorylated DCX to F-actin (Tsukada et al., 2005). Several studies have shown that Spn preferentially binds to PP1γ1 and PP1α isoforms in brain extracts (MacMillan et al., 1999; Terry-Lorenzo et al., 2002; Carmody et al., 2004). GST-Spn fusion proteins containing the PP1cbinding domain potently inhibit PP1 enzymatic activity in vitro (Allen et al., 1997; Colbran et al., 2003). However, it was recently shown that instead of inhibiting PP1c directly, Spn regulated enzymatic activity by directing its substrate specificity (Ragusa et al., 2010). Spn can associate with the tyrosine phosphatase SHP-1 and the complex modulates platelet The complex adopts a significant more extended conformation in solution than in the crystal structure. This is the result of a flexible linker (ramino acids 490-494) between the PP1c-binding and the PDZ domains. The four residue flexibility is likely important for Spn biological role (Ragusa et al., 2011). Spn also contains a single consensus sequence in PDZ, amino acids 494-585 (Allen et al., 1997). The structure of the Spn PDZ domain has been recently solved by NMR spectroscopy. The PDZ domain directly binds to carboxy-terminal peptides derived from glutamatergic AMPA and NMDA receptors (Kelker et al., 2007). Sequence analysis predicted that the carboxyterminal region of Spn (amino acids 664-814) forms 3 coiled-coil domains. Neurabins were observed as multimeric forms in vitro and in vivo. Spn and neurabin 1 homo- and hetero-dimerize via their carboxy-terminal coiled-coil domains (MacMillan et al., 1999; Oliver et al., 2002). Consensus sequences for phosphorylation by several protein kinases (PK), including cAMPdependent PK (PKA), Ca2+/calmodulin-dependent PK II (CaMKII), cyclin-dependent PK5 (Cdk5), extracellular-signal regulated PK (ERK) and protein tyrosine kinases were observed in Spn. Two major sites of phosphorylation for PKA (Ser-177 not conserved in human, and Ser-94) and two others sites for CaMKII phosphorylation (Ser-100 and Ser-116) were located within and near the F-actinbinding domain of Spn. The protein is phosphorylated in intact cells by PKA at Ser-94 and Ser-177 and by CaMKII at Ser-100 (Hsieh-Wilson et al., 2003; Grossman et al., 2004). Moreover, neurabins can be phosphorylated in vitro and in intact cells by Cdk5 on Ser-17 and ERK2 (MAPK1) on Ser-15 and Ser-205, phosphoSer-17 being abundant in neuronal cells (Futter et al., 2005). Several potential tyrosine phosphorylation sites lie within the coiled-coil regions, within a region adjacent to the PDZ domain and within the receptor-interacting domain. 3- The Spinophilin interactome Spn interactome includes cytoskeletal molecules (F-actin, doublecortin, neurabin 1, Spn), enzymes (like PP1 and CaMKII), regulator of G-protein Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 694 The tumour suppressor function of the scaffolding protein spinophilin nervous system. Spn was identified with other dendritic spines proteins as a protein partner of TRPC5 and TRPC6 channels (Goel et al., 2005). In cardiomyocytes, Spn targets PP1 to RYR2 via binding to a leucine zipper (LZ) motif of RYR2 and a LZ motif on Spn (amino acids 300-634) causing dephosphorylation and modulation of the channel activity (Marx et al., 2001). TGN38 is an integral membrane protein that constitutively cycles between the trans-Golgi network (TGN) and plasma membrane via endosomal intermediates. TGN38 directly interacts with the coiled-coil region of Spn, preferentially with the dimerized proteins (Stephens and Banting, 1999). Spn has been shown to interact with the nuclear protein ARF in mammalian cells. The amino acids sequence 605-726, of the coiled-coil region of Spn, seems to be involved and an intact ARF N-terminal region (amino acids 1-65) is necessary for this interaction (Vivo et al., 2001). activation by sequestering RGS10 and RGS18. The sequence surrounding the phosphorylation site Y398 in Spn fits a consensus ITIM sequence (I/V/L/SxY(p)xx(I/V/L) and forms a binding site for SHP1 (Ma et al., 2012). p70S6K is a mitogenactivated PK that regulates cell survival and growth. p70S6K interaction with neurabin 1 (Burnett et al., 1998) and Spn was demonstrated (Allen and Greengard, unpublished observation). The interaction implicates the PDZ domain of neurabins and the carboxyl-terminal five amino acids of the PK. CaMKII directly and indirectly associates with N- and C-terminal domains of Spn. Thus, Spn can target CaMKII to F-actin as well as target PP1 to CaMKII (Baucum et al., 2012). Regulator of G-protein signalling (RGS) proteins play a crucial role in the shutting off process of Gprotein-mediated responses (Ishii and Kurachi, 2003). Spn binds to different members of the RGS family (Wang et al., 2005; Wang et al., 2007). For example, Spn binds to through the 391-545 amino acids of the scaffolding protein and the 6-9 amino acids of the N-terminus of RGS8 (Fujii et al., 2008). Guanine nucleotide exchange factors (GEF) activate small G protein through the exchange of bound GDP for GTP. Several GEF were shown to interact with Spn. For example, Spn, through its carboxy-terminus containing the PDZ and coiledcoil domains interacts with kalirin-7, the neuronal GEF for Rac1 (Penzes et al., 2001). Spn interacts with some receptors that belong to the superfamily of GPCRs, mainly in the CNS. Using the 3i loop of the D2R, Spn has been identified as a protein that specifically associates with the receptor in rat hippocampal (Smith et al., 1999). The 3i loops of α2A-AR, α2B-AR, and α2C-AR subtypes interact also with Spn (Richman et al., 2001). More recently, it has been shown that the α1B-AR interacts with Spn in vitro (Wang et al., 2005). In the cerebellum, Spn can bind to the M1-m-AChR using the receptor binding domain of the scaffolding protein (Fujii et al., 2008). Spn can also interact with the M2- and M3-m-AchRs but the binding ability to the M3-m-AChR seems to be weaker than those to the M1- and M2-m-AChR (Wang et al., 2007; Kurogi et al., 2009). Moreover, Spn binds to the 3i loop of CCKA and CCKB receptors (Wang et al., 2007). The receptor binding domain of Spn also associates with the 3i loop and a conserved region of the C-terminal tails of δ- and µ-OR (Fourla et al., 2012). Spn also interacts with the ionotropic NMDA and AMPA-type glutamate receptors. PDZ domain directly binds to GluR2-, GluR3- (AMPA receptor) and NR1C2'-, NR2A/Band NR2C/D- (NMDA receptor) derived peptides (Kelker et al., 2007). TRPC ion channels are Ca2+ /cation selective channels that are highly expressed in the central Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Sarrouilhe D, Ladeveze V 4- Spinophilin as a tumour suppressor The Spn gene locus is located on chromosome 17 at position 17q21.33, a cytogenetic area frequently associated with microsatellite instability and loss of heterozygosity (LOH) observed in different human tumours. This region contains a relatively high density of known (such as BRCA1), putative as well as several yet-unidentified candidate tumour suppressor genes located distal to BRCA1 locus. Thus, several studies in breast and ovarian carcinomas have suggested the presence of an unknown tumour suppressor gene in the area that includes the Spn locus. However, despite these preliminary genetic correlations, no in-depth analysis of the role of Spn as a tumour suppressor has been made. The Amancio Carnero laboratory from the Instituto de Biomedicine de Sevilla, in Spain, have addressed this possibility in vitro and in vivo, in three articles published in 2011. In the first study, immunohistochemical analysis of 35 human lung tumours at different stages and of different histopathological grades showed that Spn protein is absent in 20% and reduced in another 37% of tumours, compared to normal lung tissue (MolinaPinelo et al., 2011). The loss of Spn expression correlated with a less differentiated phenotype, higher grade and poor prognosis. Lower or null levels of Spn also correlated with nuclear accumulation of p53, and so to mutated p53 or loss of its wild-type activity. Moreover, loss of Spn increased the tumourigenic properties of p53 deleted- or p53 mutated-lung tumour cells. The data of this study showed that Spn down-regulation in lung tumours contributes to carcinogenesis in the absence of p53. There are several mechanisms that might contribute to Spn down-regulation in 695 The tumour suppressor function of the scaffolding protein spinophilin regulates the expression of p14 (ARF) (Liu et al., 2012). Some members of the family of e2F transcription factors are also involved in cell cycle regulation; in particular E2F1 which expressions increase induces augmentation of ARF which can bind MDM2 and stabilize p53. In p53 (- / -) MEF, reduced levels of Spn enhanced tumorigenic potential of the cells. Indeed, inhibition of e2F by Rb being lifted, this results in cell proliferation no longer controlled by p53. Moreover, the absence of Spn contributes to genetic alterations during MEF immortalization, particularly p53 mutations. These results extend the observations made by the authors using a Spn-null mice model (Ferrer et al., 2011b). In summary, the results suggested that Spn is a new tumour suppressor acting via the regulation of pRb and which function is revealed in the absence of a functional p53 (Sarrouilhe and Ladeveze, 2012). This is, therefore, suggestive of partially redundant functions in their tumour suppression properties (Santamaría and Malumbres, 2011). The results also suggest that the specific outcome can be context-dependent. Spn loss may be beneficial by potentiating p53 in response to acute stress, and in contrast it can be deleterious under sustained mitogenic stress (Palmero, 2011). This feature is reminiscent of NIAM (Nucleolar Interaction of ARF and MDM2 protein) which acts through the same partners p53 and ARF (Tompkins et al., 2007). Another Spn-interacting molecule is DCX, an actinbinding and microtubule-binding protein that seems to be a tumour suppressor of glioma. When DCX is ectopically expressed into the DCX-deficient U87 glioma cells, there is a marked suppression of the transformed phenotype. The cells manifest a reduced rate of growth in vitro and are arrested in the G2 phase of the cell cycle. Moreover, DCXtransfected U87 glioma cells do not generate tumours in immunocompromised nude rats. In DCX-transfected U87 cells, phosphorylated DCX binds specifically to Spn and this interaction inhibits proliferation and anchorage-independent growth in glioma cells. In contrast, DCX-mediated growth repression is lost in glioma cells treated with siRNA to Spn and in HEK 293 (human embryonic kidney) Spn null cell line (Santra et al., 2006). DCX, Spn and PP1c were found in the same protein complex from mouse brain extracts (Shmueli et al., 2006). DCX-mediated growth arrest in glioma cells may be through inactivation of PP1 activity by Spn/DCX interaction in the cytosol. Inhibition of PP1 activity is involved in two mechanistic links of reduction of glioma tumour-associated progressions: firstly, catastrophe in mitotic microtubule spindle that blocks mitosis; secondly, depolymerization of actin that inhibits glioma cell invasion (Santra et al., 2009). tumours, including miRNAs overexpression. miRNA106*, targeting Spn, are overexpressed in a small subset of patients with decreased Spn levels. Overexpression of miRNA106* significantly increased the tumorigenic properties of lung tumour cells. The results suggested that miRNA106* overexpression found in a subset of lung tumours might contribute to tumorigenesis through Spn down-regulation in the absence of p53. In a second study, tumour suppression by Spn was explored in in vivo model using genetically modified mice (Ferrer et al., 2011b). Spn-null (-/-) mice displayed decreased survival, increased the number of premalignant lesions in tissues such as the mammary ducts and early appearance of spontaneous tumours, such as lymphoma, when compared to WT littermates. In another series of experiments, the presence of mutant p53 activity (p53R172H) in the mammary glands was evaluated on a Spn heterozygous (+/-) or homozygous (-/-) background in mice. An increased number of premalignant lesions and of mammary carcinomas were observed in Spn heterozygous (+/-) or homozygous (-/-) mice when compared to WT littermates. The results confirmed the functional relationship between Spn and p53 in tumorigenicity and showed that Spn loss contributes to tumour progression rather than the tumour initiation. In a third study using mouse embryonic fibroblasts (MEFs), it was suggested that Spn acts as a tumour suppressor by the regulation of the stability of PP1cα, thereby regulating its activity on pRb (the phosphorylated form of the Retinoblastoma protein). This function of PP1cα has been associated with the growth arrest response; the hypophosphorylated form of Rb protein being the most abundant when cells are delayed in their growth (Ceulemans and Bollen, 2004). The ectopic overexpression of Spn in immortalized MEF greatly reduced tumour cell growth. Moreover, the absence of Spn (Spn(-/-) MEF) down-regulated PP1α activity resulting in a high level of pRb (Ferrer et al., 2011a). High level of proproliferative phosphorylated Rb leads to e2F activation, a compensatory ARF transcription, and consequently p53 activation. As they regulate the cell cycle, p53 and ARF are both tumour suppressors, which are themselves regulated by MDM2 (Mouse double minute 2) protein shuttle between the nucleus and cytoplasm (Kamijo et al., 1998; Pomerantz et al., 1998). Moreover, Sherr et al. (2005) suggested for the first time a p53-independent pathway via the ARF sumoylation. Ha et al. (2007) described ARF as a melanoma tumour suppressor by inducing p53independent senescence. Moreover, Du et al. (2011) demonstrated the functional roles of ERK and p21 for ARF in p53-independent tumour suppression. Furthermore, in a p53-independent pathway, the over-expression of wild-type c-myc obviously up- Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Sarrouilhe D, Ladeveze V 696 The tumour suppressor function of the scaffolding protein spinophilin Sarrouilhe D, Ladeveze V Figure 3. Cellular cycle regulation by spinophilin. A. In normal cells, the presence of nuclear p53 and Spn proteins regulates cell cycle. The binding of PP1ca to Spn allows dephosphorylation of pRb, which inhibits E2F1 and thus the proliferation. Furthermore both tumour suppressors (p53 and ARF) regulate the cell cycle. The nucleolar ARF is also a partner of Spn, and regulates the cell cycle via Mdm2 and E2F1. B. In the case of colorectal carcinomas, Spn play a role in regulation of cell cycle via a p53/ARF independent pathway. One hypothesis suggested by the team of Amancio Carnero is that the Ras/Raf pathway could be implicated (Estevez-Garcia et al., 2013). This cytoplasmic pathway could be regulated by cytoplasmic Spn. K-Ras: GTPase, oncogene; B-Raf: serine/threonine protein kinase, proto-oncogene; Mek: tyrosine/threonine kinase (Mapk kinase); Mapk: mitogen-activated protein kinase. diagnosed after the 10-year follow-up in 85.2% cases with Spn low expression and 60.9% with Spn high expression. Death occurred in 76.5% cases with Spn low expression and in 56.5% cases with Spn high expression. Overall, low Spn expression is a factor for poor prognosis in hepatocellular carcinoma. In vitro experiments (human hepatoma cell line HepG2) and in vivo observations (Ki67positive tumour cells) showed that reduced Spn expression significantly correlated with a higher proliferation of liver cancer cells (Aigelsreiter et al., 2013). In the second study, the role of Spn was explored in colorectal carcinoma, in which a number of chromosomal regions are altered (Fearon, 2011). Among them, the 17q21 is lost in a high percentage of this carcinoma (Garcia-Patiño et al., 1998). Quantitative RT-PCR analysis showed that approximately 25% of colorectal carcinoma tumours had a greater than 50% decrease in Spn mRNA levels compared with normal colonic tissue. A tissue array of human colorectal carcinomas was generated to confirm this result by exploring the presence of Spn protein. 70% of colorectal carcinomas displayed high Spn levels (similar to the values observed in normal tissue), 20% showed intermediate levels and 10% showed no expression of Spn. Moreover, Spn down-regulation correlated with a more aggressive histologic phenotype (higher Ki67-positive tumour cells) and was associated with faster relapse and poorer survival in patients with advanced stages of colorectal carcinoma. The data also suggested that Spn loss induced a chemoresistance in patients with advanced stages of colorectal carcinoma that had received adjuvant fluoropyrimidine chemotherapy Moreover, double transfection with DCX and Spn reduced self-renewal in brain tumour stem cells via incomplete cell cycle endomitosis (Santra et al., 2011). But, is there relevance for Spn as a prognostic marker in patients with cancer? Spn is absent in 20% and reduced in another 37% of human lung tumors (Molina-Pinelo et al., 2011). A further analysis of Spn in human tumours shows that Spn mRNA is lost in a percentage of renal carcinomas and lung adenocarcinomas. A clear down-regulation of Spn was found in tumoral samples of the CNS (oligodendrogliomas, anaplastic astrocytomas, glioblastomas) when compared to normal nervous samples. Furthermore, lower levels of Spn mRNA correlate with higher grade of ovarian carcinoma and chronic myelogenous leukemia (Carnero, 2012). Two articles published in spring 2013 associated Spn loss with poor patient prognosis in patients with carcinoma (Sarrouilhe, 2014). The 17q chromosomal region is commonly impaired in hepatocellular carcinoma (Furge et al., 2005). In the first study, complete loss of Spn immunoreactivity was found in 42.3% hepatocellular carcinoma and reduced levels were found in additional 35.6% cases. Quantitative RT-PCR analysis confirmed in 70% cases a significant reduced Spn mRNA expression in tumour tissue compared with the corresponding non-neoplastic tissue. miRNA106*, targeting Spn in lung tumours, could not be detected in any of the hepatocellular carcinoma samples. Moreover, no correlations could be found for the number of Spn-positive tumour cells and p53 or ARF staining. These results suggested a p53-independent tumorigenic role of Spn in hepatocellular carcinoma. Disease recurrence was Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 697 The tumour suppressor function of the scaffolding protein spinophilin Sabatini DM, Snyder SH. Neurabin is a synaptic protein linking p70 S6 kinase and the neuronal cytoskeleton. Proc Natl Acad Sci U S A. 1998 Jul 7;95(14):8351-6 following surgical resection. Therefore, the identification of the levels of Spn in advanced stages of colorectal biopsies has prognostic and predictive value and might contribute to select patients who could or could not benefit from current chemotherapy. In vitro and in vivo experiments showed no functional relationship between Spn levels and the presence or absence of mutated p53 in colon cancer. The authors proposed that this correlation is dependent on the molecular context of the tumour cell (Estevez-Garcia et al., 2013). 5- Discussion and perspectives We are still only at the early stage in unravelling the function of Spn in cell cycle regulation. Overall, the different studies on the tumour suppressor function of Spn show two pathways of cell cycle regulation by Spn. The first model is a pathway dependent of p53 and ARF. This pathway was previously described in several articles where Spn interacts with different partners localized in the nucleus (Figure 3A). The second is a pathway independent of both molecules. As Spn is ubiquitously expressed in the cell, the first model highlights the nuclear localization of Spn and its interaction with other nuclear proteins. The second model, more hypothetical, underlines the possibility that Spn could interact with cytoplasmic partners. The studies made on colorectal carcinomas show that Spn could play a role in a pathway independent of p53/ARF. One hypothesis is that the Ras/Raf pathway and more precisely K-Ras/B-Raf is implicated. This pathway, via Mek (tyrosine/threonine kinase) and Mapk (mitogen activated protein kinase) induces transcription factors and proliferation survical (Figure 3B). Further studies are needed to elucidate the underlying mechanisms linking Spn to carcinomas and expand the prognostic and predictive value of the Spn expression level to other types of cancer. 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Bull Cancer. 2014 Jan 1;101(1):5-6 Santamaría D, Malumbres M. Tumor suppression by Spinophilin. Cell Cycle. 2011 Sep 1;10(17):2831-2 This article should be referenced as such: Santra M, Santra S, Buller B, Santra K, Nallani A, Chopp M. Effect of doublecortin on self-renewal and differentiation in brain tumor stem cells. Cancer Sci. 2011 Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Sarrouilhe D, Ladeveze V Sarrouilhe D, Ladeveze V. The tumour suppressor function of the scaffolding protein spinophilin. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):691-700. 700 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Case Report Section Paper co-edited with the European LeukemiaNet Translocation t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia patient Adriana Zamecnikova, Soad Al Bahar, Ramesh Pandita Kuwait Cancer Control Center, Department of Hematology, Laboratory of Cancer Genetics, Kuwait (AZ, SA, RP) Published in Atlas Database: February 2014 Online updated version : http://AtlasGeneticsOncology.org/Reports/t0506q33q23ZamecID100076.html DOI: 10.4267/2042/54042 This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 2.0 France Licence. © 2014 Atlas of Genetics and Cytogenetics in Oncology and Haematology Immunophenotype Positive for CD13, CD15, CD117, CD33, MPO, CD45, HLDR and dim CD34 (27%) Diagnosis Acute myelomonocytic leukemia Abstract Case report and literature review on translocation t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia patient. Clinics Survival Age and sex 68 years old female patient. Previous history No preleukemia, no previous malignancy, no inborn condition of note, no main items. Organomegaly No hepatomegaly, no splenomegaly, no enlarged lymph nodes, no central nervous system involvement. Date of diagnosis: 03-2013 Treatment: Chemotherapy (Daunorubicin & Cytarabine combination therapy; consolidation with high dose Ara-C) Complete remission: no Treatment related death: no Relapse: yes Phenotype at relapse: Acute myelomonocytic leukemia Status: Lost Last follow up: 11-2013 Survival: 8 months Blood WBC: 104 X 109/l HB: 7.8g/dl Platelets: 57 X 109/l Blasts: 84% Bone marrow: Hypercellular marrow with 87% blasts which were PAS diffuse granular positive and SBB (Sudan Black B) positive. Karyotype Sample: Bone marrow, blood Culture time: 24h Banding: G-banding Results 46,XX,t(5;6)(q33-34;q23)[25] Karyotype at Relapse 46,XX,t(5;6)(q33-34;q23)[1]/46,XX,t(5;6)(q3334;q23),t(7;10)(p22;q23)[19] Cyto-Pathology Classification Cytology Acute myelomonocytic leukemia Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 701 Translocation t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia patient Zamecnikova A, et al. Figure 1. A. Karyotype from the time of diagnosis showing the chromosomal translocation t(5;6)(q33-34;q23). B. Fluorescence in situ hybridization studies (FISH) with XL 6q21/6q23 (Metasystem, Germany) probe showing red and green signals on both, normal and der(6) chromosomes. C. Applying the XL PDGFR probe (Metasystem, Germany) showed normal signal pattern on both normal and der(5) chromosomes, indicating that PDGFR located on 5q32-33 is not involved in the translocation. D. Hybridization with whole chromosome 6 probe (Metasystem, Germany) showing translocation of chromosome 6 sequences to der(5) chromosome. ALL and an associated myeloproliferative neoplasm and C6ORF204/PDGFRB fusion (Chmielecki et al 2012). While in this case, the chromosomal translocation appeared to be morphologically identical to our t(5;6)(q33-34;q23), in our patient PDGFRB (5q3233) is not rearranged and MYB (6q23)is not translocated to chromosome 5 as in a previously described case. Due to the availability of tyrosine kinase inhibitors for PDGFRB rearranged disorders, our findings emphasize the importance of FISH in precise characterizing of chromosome rearrangements with 5q33-34 breakpoints, especially in suboptimal preparations. Other molecular cytogenetics technics Fluorescence in situ hybridization (FISH) with LSI AML1-ETO, LSI MLL, LSI CBFB/inv(16), LSI EGRI/5q31 (Abbott Molecular, Downers Grove, IL) and XL 6q21/6q23, XL PDGFR, whole chromosome 6 probe Metasystem, Germany). Other molecular cytogenetics results Normal signal patterns for LSI AML1-ETO, LSI MLL, LSI CBFB/inv(16), LSI EGRI/5q31, XL 6q21/6q23 and XL PDGFR probes. Comments Chromosomal translocations involving 5q33 and 6q23 have been reported in only one patient with T- Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 702 Translocation t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia patient Zamecnikova A, et al. Figure 2. A. Karyotype from blood cell from the time of relapse showing the t(5;6)(q33-34;q23) and a new anomaly t(7;10)(p22;q23). B. Partial karyotypes from blood and bone marrow showing the t(5;6)(q33-34;q23). Chromosomes Cancer. 2012 Jan;51(1):54-65 References This article should be referenced as such: Chmielecki J, Peifer M, Viale A, Hutchinson K, Giltnane J, Socci ND, Hollis CJ, Dean RS, Yenamandra A, Jagasia M, Kim AS, Davé UP, Thomas RK, Pao W. Systematic screen for tyrosine kinase rearrangements identifies a novel C6orf204-PDGFRB fusion in a patient with recurrent T-ALL and an associated myeloproliferative neoplasm. Genes Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) Zamecnikova A, Al Bahar S, Pandita R. Translocation t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia patient. Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9):701-703. 703 Atlas of Genetics and Cytogenetics in Oncology and Haematology INIST-CNRS OPEN ACCESS JOURNAL Instructions to Authors Manuscripts submitted to the Atlas must be submitted solely to the Atlas. Iconography is most welcome: there is no space restriction. The Atlas publishes "cards", "deep insights", "case reports", and "educational items". Cards are structured review articles. 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See also: "Uniform Requirements for Manuscripts Submitted to Biomedical Journals: Writing and Editing for Biomedical Publication - Updated October 2004": http://www.icmje.org. http://AtlasGeneticsOncology.org © ATLAS - ISSN 1768-3262 Translocation t(5;6)(q33-34;q23) in an acute myelomonocytic leukemia patient Atlas Genet Cytogenet Oncol Haematol. 2014; 18(9) 705 Zamecnikova A, et al.