Domain strcture of SUMO ligase PIAS1 ant ispecific interaction of its target protein p53
| Project/Area Number |
16590034
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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 |
Physical pharmacy
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| Research Institution | Tokyo University of Pharmacy and Life Sciences |
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Principal Investigator |
SHINDO Heisaburo Tokyo University of Pharmacy and Life Sciences, School of Pharmacy, Professor, 薬学部, 教授 (80138966)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2004: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | SUMO ligase / PIAS1 / Tumor seppresor p53 / NMR structure / transcription factor Lef-1 / PIASy / Siz1 / SUMOリガーゼ / SAPドメイン / DNA結合蛋白質 |
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| Research Abstract |
Recently, many kinds of ubiquitin-like proteins are discovered and the roles of such post-modifications are paid strong attention. SUMO (small ubiquitin like modifier) is one of them and now it becomes clear that sumoylation plays important roles in localization to nuclear, regulation of transcription, and moreover in the process of segregation of chromosome. In this research project, we want to clarify (1)determination of 3D structure of PIAS1, (2)recognition mechanism of tumor suppressor p53 and transcription regulators such as Lef-1 and c-jun by PIAS1, (3)Structure determination of other members of PIAS family such as PIASy and Siz1.
In order to identify binding sites of p53 and PIAS1, we have prepared a series of deletion mutants of PIAS1, investigated specific interaction of p53 and the N-terminal domain of PIAS1 using GST-pull down assay. The results showed that the N-terminal domain interacts p53, and that the minimum length of PIAS1 for the binding is the amino acid segment (1-65). This N-terminal domain was also shown to interact with DNA as well as transcription factor Lef-1 HMG box. DNA was recognized by a2- and a3- helix of N-terminal domain of PIAS1, but did not bind c-jun.
We have determined 3D structure of the N-terminal domain by multi-dimensional NMR, which was an unusual four-helix bundle. Now, we have constructed the N-terminal domain (1-72) of subtype of PIAS1, PIASy, and the domain (1-111) of yeast SUMO ligase Siz1. The recombinant proteins were overexpressed and purified. Now, we are measuring a series of multi-dimensional NMR for structure determination.
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Report
(3 results)
Research Products
(7 results)
