Download Ligand Identification for Orphan Nuclear Receptors TLX and PNR.

Title of Research Project: Ligand Identification for Orphan Nuclear Receptors TLX and
PNR(2 years in U.S.A.+2 years in NUS graduate programme)
Name of Supervisors:
Professor E. L. Yong (Head, Dept of Obstetrics & Gynaecology ),
Distinguished Investigator H. Eric Xu (Van Andel Research Institute, U.S.A.)
Contact Details: [email protected], [email protected]
Short Description
Nuclear hormone receptors are key DNA-binding and ligand-inducible transcriptional factors that regulate neural
development, sexual function, reproduction and metabolism. There are 48 nuclear hormone receptors in the human
genome including classic steroid hormone receptors, thyroid hormone receptor and vitamin A and vitamin D receptors
with known ligands. There are also a few orphan nuclear receptors for which their cognate ligands are not known.
Since ligand regulated transcription activity is a key aspect of nuclear receptor function, identification of ligand for
orphan nuclear receptors is critical for understanding the biology of these receptors. This project is to identify the
ligand for two orphan receptors TLX and PNR, which play central role in the development of eye and central nerve
system. To gain insights into the transcriptional regulation of the TLX/PNR receptor after knowing their ligands,
chromatin immunoprecipitation coupled with ultra-high-throughput DNA sequencing (ChIP-seq, done in Biopolis) will
be used to map the binding locations of TLX/PNR in the genome. We will be able to map all TLX/PNR and RNA
polymerase II binding sites on a genome-wide scale, in vivo, to identify the authentic cis binding sites and target genes
in a Y79 cell line. Combining this unique resource with gene expression data from microarray, we will demonstrate
distinct temporal mechanisms of the novel TLX/PNR ligand in the mediation of gene regulation in both suppressed and
activated genes.
This is a multidisciplinary project under a research collaboration agreement between NUS and Van Andel Research
Institute, Michigan, U.S.A. which allows the student to participate a “2+2” graduate programme and have access to
many facilities in different laboratories required for completing the project. Furthermore, proper training will be
provided from experienced lab technicians/senior research fellows/associate professor to master the core techniques
including recombinant protein expression on mammalian system and gene transcription regulation by realtime RTPCR, ForteBio’s Octet biosensor and Amplified Luminescent Proximity Homogeneous Assay to understand
biomolecular interaction, protein large scale expression and affinity purification by FPLC, protein crystallization using
Phoenix crystallization robot which are required for the study. Detail on this project can be acquired by sending
email request to: Dr. Li Jun (Tel: 65165168, [email protected] or [email protected]). Further details on the PIs can be
found at: http://www.nus.edu.sg/ngs/supervisors_cv/yong_eu_leong.pdf
http://www.vai.org/Research/Labs/StructuralSciences.aspx
Recent Relevant Publications:
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Schopfer FJ, Cole MP, Groeger A, Chen CS, Woodcock SR, Khoo N, Golin-Bisello F, Motanya UN, Rudolph TK,
Rudolph V, Bonacci G, Baker PRS, Batthyany CI, Xu HE, Y. Chen YE, Hallis TM, and Freeman BA (2008)
Covalent Peroxisome Proliferator-Activated Receptor Binding by Nitro-Fatty Acids: Endogenous Ligands Act as
Selective Modulators. Nature Medicine, (in review)
Xu HE and Li Y (2008) Ligand-dependent and –independent regulation of PPAR and Orphan Nuclear receptors.
Science Signaling 1(48):pe52 (Cover Article)
Kruse SW, Suino-Powell K, Zhou E, Krestchman JE, Reynolds R, Vonrhein C, Xu Y, Wang L, Tsai SY, Tsai MJ,
Xu HE (2008) Identification of COUP-TFII Orphan Nuclear Receptor as a Retinoic Acid Activated Receptor.
PLOS Biology, 6(9):e227.
Yong EL, Li J, Liu MH. (2008) Single gene contributions: genetic variants of peroxisome proliferator-activated
receptor (isoforms alpha, beta/delta and gamma) and mechanisms of dyslipidemias. Curr Opin Lipidol. 19(2):10612.
Liu MH, Li J, Shen P, Husna B, Tai ES, Yong EL.(2008) A natural polymorphism in peroxisome proliferatoractivated receptor-alpha hinge region attenuates transcription due to defective release of nuclear receptor
corepressor from chromatin. Mol Endocrinol. 22(5):1078-92.