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2013년 한국분자세포생물학회 식물분자생물학분과 심포지엄 및 제1회 식물후성유전체학 심포지엄 “Epigenomics in Plants 2013” 주관: 한국분자세포생물학회 및 차세대유전체연구사업단 일시 및 장소: 2013년 11월 11일, 서울대학교 25-1동 국제회의실 13:30 ~ 14:00: 등록 및 한국식물후성유전학연구회 회원 접수 14:00 ~ 14:10: 인사의 말씀 (차세대유전체사업단장 박범석, 식물분자생물학분과장 이일하) I. 한국분자세포생물학회 식물분자생물학분과 주관 세션 (좌장: 노유선) 14:10 ~ 14:35 강훈승 (전남대 교수): RNA-binding proteins, RNA chaperones, and RNA metabolism in plant development and stress responses 14:35 ~ 15:00 최연희 (서울대 교수): MOS7, an Arabidopsis Nup88 homolog, is crucial for gamete formation 15:00 ~ 15:25 이병하 (서강대 교수): Splicing-involved RNA biology 15:25 ~ 15:40 석혜연 (부산대 박사과정): EMF1 interacts with EIP1, EIP6 or EIP9 involved in the regulation of flowering time in Arabidopsis 15:40 ~ 16:00 Coffee Time II. 차세대유전체연구사업단 주관 세션 (좌장: 신찬석) 16:00 ~ 16:25 최상봉 (명지대 교수): Puf RNA-binding proteins and RNA decay 16:25 ~ 16:50 허진회 (서울대 교수): Nonadditive transcriptional regulation in xBrassicoraphanus derived from intergeneric hybridization between Brassica rapa and Raphanus sativus 16:50 ~ 17:15 허재복 (동아대 교수): Long non-coding RNA-mediated epigenetic repression of flowering pathway genes in Arabidopsis 17:15 ~ 17:30 정영민 (가톨릭대 연구교수): Distinctive roles of two JmjC domain-containing proteins in the repression of FLC in Arabidopsis 17:30 ~ 17:45 Closing Announcement & Photo Time (※) 본 심포지엄에서는 한국식물후성유전학연구회(회장 노유선, 간사 신찬석)의 창립을 기하여 정회원(박사급) 및 준회원(연구원 및 대학원생) 등록을 진행합니다(회비 없음). 후성유전학에 관심 있는 모든 분들의 적극적 참여를 부탁 드립니다. RNA-binding Proteins, RNA Chaperones, and RNA Metabolism in Plant Development and Stress Responses Hunseung Kang Department of Plant Biotechnology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju, 500-757 Korea Posttranscriptional RNA metabolism is recognized as a key regulatory process in the growth, development, and stress response of plants, and a variety of RNA-binding proteins (RBPs) have been demonstrated to perform central roles in these cellular processes. In particular, certain RBP family members play their roles as RNA chaperones that aid in proper RNA folding via structural rearrangement during RNA metabolism. During the last years, we have extensively investigated the functional roles of diverse RBP family members in the process of plant growth and development as well as the stress response of plants. RBPs unique to minor spliceosome complex and is involved in the splicing of U-12 type minor introns, glycine-rich and zinc finger-containing RBPs, cold shock proteins, and RNA helicases have been extensively studied for their roles in plant growth and stress responses. In addition, functional analysis of chloroplast-targeted nuclear-encoded RBPs showed that they play crucial roles for the gene expression, biogenesis, and function of chloroplasts under normal and stress conditions. All of these results point to the importance of posttranscriptional regulation of RNA metabolism in plants. In this talk, discussed will be recent progress and novel findings as well as remaining challenging questions on the functional roles of RBPs and RNA chaperones in plant growth, development, and stress responses. MOS7, an Arabidopsis Nup88 Homolog, Is Crucial for Gamete Formation Geun-Tae Park and Yeonhee Choi Department of Biological Sciences, Seoul National University, Seoul 151-747, Korea One of the most striking differences between the developmental approaches of plants and animals is that the several mitotic divisions intervene between meiosis and gamete formation. Mature plant gametes are produced after a couple of successive mitotic divisions in haploid cells that exist only in flowering plants and following cytokinesis are critical. However, the information and mechanisms of genes which mediate these major events during both gametophyte developments are still largely unknown. Here, we show that MOS7 (Modifier Of Snc1,7) that is homologous to human and Drosophila Nup88 nucleoporin (NUP) is critical for male and female gamete formation as well as embryo development. Haplo-insufficient mos7-5 allele in megaspore mother cell (MMC) and pollen mother cell (PMC) resulted in delayed phenotypic defects after meiosis, during subsequent mitotic divisions. MOS7 is expressed in various tissues and cells including MMC and PMC. No homozygous mos7-5 mutant was obtained, indicating an embryo lethality. mos7-5 mutant showed defects in cortical microtubule in microspore and in CenH3-GFP incorporation during pollen mitosis I (PMI). Mitotic spindle assembly, spindle attachment to the kinetochore as well as fragmoplast and cell plate formation were also defected during gametogenesis in mos7-5 mutant. Yeast two hybrid screen identified dynein and kinectin which are microtubule associated proteins (MAPs) as MOS7-interacting partners. Taken together, our data suggest that MOS7 plays an important role in microtubule dynamics possibly together with MAPs during gametogenesis. Splicing-involved RNA Biology Byeong-ha Lee Department of Life Science, Sogang University, Seoul 121-742, Korea Arabidopsis STABILIZED1 (STA1) encodes a protein that is homologous to human U5 snRNP-associated 102-kDa protein (PRPF6), and the yeast pre-mRNA splicing factors, PRP1p (fission yeast) and Prp6p (budding yeast), and is important in pre-mRNA splici ng and mRNA stability. The pleiotropic defects of development, chilling sensitivity an d hypersensitivity to ABA are observed in sta1-1, a weak allele of sta1. sta1-1, show ing enhanced luminescence under cold stress, was originally isolated from a mutant p ool generated with the bioluminescent plant harboring the stress-inducible RD29A prom oter-driven luciferase gene (RD29A-LUC). Some developmental defects found in sta11 resembled those found in miRNA biogenesis mutants such as hyponastic leaves1-1 ( hyl1-1) and serrate-1 (se-1). Similar to these miRNA biogenesis mutants, sta1-1 accu mulated significantly lower levels of mature miRNAs and concurrently higher levels o f pri-miRNAs than wild type. The dramatic reductions of mature miRNAs were associ ated with the accumulation of their target gene transcripts and the corresponding devel opmental defects. The reduction of miRNA accumulation in sta1-1 appeared to be bec ause of the sta1-1 decfects in splicing of intron-containing pri-miRNAs. In addition, st a1-1 decreased transcript levels of DICER-LIKE1 (DCL1) gene. These results suggest that STA1 is involved in miRNA biogenesis directly by functioning in pri-miRNA spli cing and indirectly by modulating the DCL1 transcript level. Our efforts and recent fi ndings to establish STA1-mediated RNA metabolism will be presented and discussed. EMF1 Interacts with EIP1, EIP6 or EIP9 Involved in the Regulation of Flowering Time in Arabidopsis Hye-Yeon Seok, Hee-Yeon Park, Sun-Young Lee, Sun-Ho Kim, Vaishali N. Tarte, Z. Renee Sung and Yong-Hwan Moon Department of Molecular Biology, Pusan National University, Busan 609-735, Korea Flowering time of higher plants is finely regulated by floral inducers and floral repressors. EMBRYONIC FLOWER (EMF) 1 gene is necessary for the maintenance of vegetative development and functions as floral repressor. To investigate the molecular mechanism of EMF1-mediated plant development, we screened EMF1-interacting proteins and identified three EMF1-Interacting Proteins (EIPs), EIP1, 6 and 9, predicted to encode a WNK kinase, a B-box zinc-finger protein and a DnaJ-domain protein, respectively. All of heterodimers between EMF1 and EIP1, 6 or 9 were localized in the nucleus. The middle and the C-terminal regions of EMF1 played an important role in the interaction with EIP proteins. The Cterminal region of EIP1, including PWNK motif, and the N-terminal region of EIP6, including B-box zinc finger, were involved in the interaction with EMF1. In contrast, the entire ORF of EIP9 was necessary for the interaction with EMF1. Interestingly, although EMF1 did not directly interact with EMF2, EMF1 interacted with EMF2 via EIP9, forming a ternary protein complex of EMF1-EIP9-EMF2 and showed granule-like dispersion patterns in the nucleus. The expression patterns of EIP1, 6 and 9 were similar to that of EMF1 during vegetative development. In mature plants, the transcripts of EIP1, 6 and 9 were predominantly expressed in leaves and inflorescence stems while lowly expressed in floral organs. In addition, eip1, eip6 and eip9 flowered earlier compared to wild type, and the transcript levels of FT, PI and AG significantly increased while SOC1 and CO transcripts slightly increased in the mutants. On the contrary, EIP1-, 6- and 9-overexpressing transgenic plants showed delaying flowering with decreased levels of FT, SOC1, PI and AG. Taken together, our results suggest that EMF1 interacts with EIP1, 6 and 9 during vegetative development to regulate flowering time in Arabidopsis. Puf RNA-Binding Proteins and RNA Decay Hyung-Sae Kim, Nazia Abbasi and Sang-Bong Choi Division of Bioscience and Bioinformatics, Myongji University, Yongin, Kyunggi 449-718, Korea Post-transcriptional control including mRNA transport, stability, and translation plays a crucial role in plant growth and development. Most of these processes are inevitably achieved either directly or indirectly by RNA-binding proteins (RBPs). Although Pumilio proteins have been characterized in many eukaryotes, their roles in plants are largely unknown. Plants have more Pumilios than any other organisms, thus Arabidopsis has 25 and rice has 20. One of the Arabidopsis Pumilios, APUM23, participates in the degradation of poly(A) 18S and 5.8S rRNA by-products in nucleolus. Mutant lacking APUM23 activity showed slow growth and abnormal growth patterning. The other nucleolar Pumilio, APUM24, is required for coordinated process of female gametogenesis and embryogenesis. The apum24 mutant defects in embryo sac development followed by developmental arrest of embryos at various stages, ranging from zygote to late globular stages. APUM24 plays an important role in integrating the role of ribosome biogenesis with the proper cell cycle progression. Among other Pumilios, APUM2 colocalizes with processing-body markers, indicating its role on RNA homeostasis. APUM2 appears to be involved in the timely mRNA decay of seed storage protein genes during early germination stage. APUM1 and APUM3 have similar primary structure with APUM1, suggesting its role on seed germination. [Supported by grants from Next-Generation BioGreen21 Program, RDA (No. PJ008206)] Nonadditive Transcriptional Regulation in xBrassicoraphanus Derived from Intergeneric Hybridization between Brassica rapa and Raphanus sativus Jin Hoe Huh Department of Plant Science, Seoul National University, Seoul 151-921, Korea Hybrids and polyploids occur in many flowering plants. In particular, hybridization between different species or genus may produce novel plant species, which is best exemplified in the Brassica family. However, many interspecific or intergeneric hybrids have reduced viability and fertility due probably to genome incompatibilities caused by interactions between genes that are functionally diverged in the respective hybridizing species. xBrassicoraphanus, also known as Baemoochae, is a newly synthesized intergeneric allotetraploid between Chinese cabbage (Brassica rapa L.) and radish (Raphanus sativus L.). Unlike most interspecific or intergeneric allopolyploids, xBrassicoraphanus is a rarely successful polyploid displaying intermediate phenotypes for many traits. Their stable inheritance and great fertility indicate that genome incompatibility was overcome after hybridization partly by alleviating a ‘transcriptome shock’ caused by two diverged genomes. Upon hybridization, many of orthologous genes become homoeologous gene pairs in the same nucleus and they are expected to be under control of common trans-acting regulators. Also, many transcription factors and genes involved in signal transduction may reconstitute the signaling networks. In this study, we obtained the RNA-seq data from xBrassicoraphanus and its respective parents Brassica rapa and Raphanus sativus using Illumina HiSeq 2000. We performed de novo transcriptome assembly for xBrassicoraphanus and identified homoeologous genes by comparing with the two parental transcriptome data. We found that many of homoeologous genes in xBrassicoraphanus were differentially expressed compared to their hybrid parents. Interestingly, a significant number of genes were downregulated in a non-additive fashion deviating from mid-parental value in xBrassicoraphanus. This suggests that global transcriptional suppression for two parental genomes is one of the mechanisms to stabilize the gene expression network in the hybrid between the distantly related species. Long Non-coding RNA-mediated Epigenetic Repression of Flowering Pathway Genes in Arabidopsis Jae Bok Heo Department of Molecular Biotechnology, Dong-A University, Busan, 604-714, Korea Many eukaryotes, including plants, produce a number of long noncoding RNAs (lncRNAs). Growing number of lncRNAs are being reported to have regulatory roles in diverse developmental processes. In Arabidopsis, vernalization results in stable epigenetic repression of a potent floral repressor, FLOWERING LOCUS C (FLC). Increased enrichment of Polycomb Repressive Complex 2 (PRC2) and trimethylated Histone H3 Lys 27 (H3K27me3) at FLC chromatin is necessary for the stable maintenance of FLC repression by vernalization. A long intronic noncoding RNA (termed as COLDAIR) is required for the vernalizationmediated epigenetic repression of FLC. COLDAIR physically associates with a component of PRC2 and targets PRC2 to FLC. COLDAIR is required for establishing stable repressive chromatin at FLC through its interaction with PRC2. In addition, floral integrator genes are targets of PRC2 complex, resulting in delayed flowering time through repression mechanism of PRC2 complex. Recently another long non-coding RNA was isolated from floral integrator gene and characterized the function of this long non coding RNA. Distinctive Roles of Two JmjC Domain-containing Proteins in the Repression of FLC in Arabidopsis Young-Min Jeong1,2,†, Jae-Young Yun2, Jung-Nam Cho1, Richard M. Amasino2 and Yoo-Sun Noh1 1 School of Biological Sciences, Seoul National University, Seoul 151-747, Korea; 2 Department of Biochemistry, University of Wisconsin-Madison, Madison, WI 53706 USA; † Current address: Department of Biology, The Catholic University of Korea, Bucheon, 420743, Korea Histone modifications often impose an epigenetic layer of regulation on eukaryotic gene expression. Methylation of histone, which is regulated by diverse histone methyltransferases and demethylases, is one of the important histone modifications. Jumonji C (JmjC) domaincontaining proteins have been reported as histone demethylases, and among the twenty-one JmjC protein members of Arabidopsis only a few have been functionally characterized. Here we report that mutations in Arabidopsis thaliana Jumonji 12 (AtJmj12) result in late flowering caused by increased expression of FLOWERING LOCUS C (FLC), the major floral repressor in Arabidopsis. The late-flowering of atjmj12 was suppressed by an flc null mutation and by vernalization, indicating that AtJmj12 affects flowering through FLC and a member of the autonomous pathway. Chromatin immunoprecipitation assays showed that AtJmj12:FLAG fusion protein associate directly with FLC chromatin. The double mutants between atjmj12 and relative of early flowering 6 (ref6), another JmjC domain-containing FLC repressor, flowered drastically later than either of the single mutants and possessed increased level of FLC expression. However, mutations in atjmj12 and ref6 caused distinct patterns within FLC chromatin, indicating that AtJmj12 and REF6 repress FLC expression through distinct mechanisms.