2007 Fiscal Year Final Research Report Summary
Comprehensive identification of nuclear receptor-regulating proteins usinghigh luminescent YFP-fusion cDNA library
| Project/Area Number |
18591027
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Endocrinology
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| Research Institution | Kyushu University |
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Principal Investigator |
KAWATE Hisaya Kyushu University, Hospital, Assistant Professor (20336027)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAYANAGI Ryoichi Kyushu Unyversity, Graduate School of Medical Sciences, Professor (30154917)
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| Project Period (FY) |
2006 – 2007
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| Keywords | Nuclear receptor / Gene / Confocal microscope / Hormone |
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| Research Abstract |
1. Examination of the significance of nuclear foci formation of steroid hormone receptors We hound that ligand-dependent foci formation of steroid hormone receptors does not precede the transcriptional activation and depends on the relative expression ratio of coactivators and corepressors.
2.Domain analysis of transcription compressor N-CoR We generated various N-CoR deletion mutants and hound that the central portion (1133-1798 amino acid) of the protein was mostly responsible for both the interaction with steroid hormone receptors and the repression of their activities.
3.Mutual transactivational repression of Runx2 the androgen receptor (AR) Tb examine the role of steroid hormone receptors on bone metabolism, we analized mutual action between steroid hormone receptors and Runx2, which is a key regulator for osteoblast diffrentiation and proliferation. Our experimental results suggest that both AR and Runx2 repress the transcriptional function of the other protein by extracting it from its original compartment.
4.Construction of high luminescentYFP-cDNAlibrary and introdution into culture cells Messenger RNA was extracted from HeLa cells and cDNA was synthesized by reverse transcriptase and then fused to high luminescent YFP vector to make fusion library. This cDNA library was cotransfected with AR-expression plasmid into COS-7 cells. After the ligand treatment, we tried to collect cells showing specific nuclear patterns similar to those of AR. However, we failed to pick up such kind of cells because cells showing foci-like pattern was very rare.
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