Domain strcture of SUMO ligase PIAS1 ant ispecific interaction of its target protein p53

SUMO连接酶PIAS1的结构域及其靶蛋白p53的特异性相互作用

基本信息

批准号：
16590034
负责人：
SHINDO Heisaburo
金额：
$ 2.24万
依托单位：
Tokyo University of Pharmacy and Life Sciences
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590034/
关键词：
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

项目摘要

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.

最近，发现了许多类型的泛素样蛋白质，并引起了这种变化后的作用。 Sumo（像修饰剂一样小的泛素）是其中之一，现在很明显，Sumoylation在定位，转录调节以及在染色体隔离过程中起着重要作用。 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,使用GST-PULL向下测定法研究了p53和PIAS1的N末端结构域的特定相互作用。结果表明，N末端结构域相互作用p53，并且结合的PIAS1最小长度是氨基酸段（1-65）。该N末端结构域还显示与DNA以及转录因子LEF-1 HMG盒相互作用。 DNA由PIAS1的N末端结构域的A2-和A3-螺旋识别，但没有结合C-Jun。我们通过多维NMR确定了N末端结构域的3D结构，这是一个不寻常的四螺旋束。现在，我们构建了PIAS1，PIASY和PIASY和酵母Sumo连接酶Siz1的域子类型的N末端结构域（1-72）。重组蛋白过表达并纯化。现在，我们正在测量一系列的多维NMR来确定结构。

项目成果

期刊论文数量（14）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Hydrogen-bond patterns in the hydration structure of a protein

DOI：
10.1016/j.cplett.2004.11.071
发表时间：
2005-01
期刊：
Chemical Physics Letters
影响因子：
2.8
作者：
T. Yokomizo;M. Nakasako;T. Yamazaki;H. Shindo;J. Higo
通讯作者：
T. Yokomizo;M. Nakasako;T. Yamazaki;H. Shindo;J. Higo

Mitochondria-specific RNA-modifying Enzymes Respongible for the Biosynthesis of the Wobble Base in Mitochondrial tRNAs

线粒体特异性 RNA 修饰酶负责线粒体 tRNA 中摆动碱基的生物合成

DOI：
发表时间：
2005
期刊：
J. Biol. Chem 280
影响因子：
0
作者：
N.Umeda;T.Suzuki;M.Yukawa;Y.Ohya;H.Shindo;K.Watanabe;T.Suzuki
通讯作者：
T.Suzuki

NMR structure of the N-termnal domain of SUMO ligase PIAS1 and its interaction with tumor supressor p53 and A/T-rich DNA oligomers

SUMO连接酶PIAS1 N端结构域的NMR结构及其与肿瘤抑制因子p53和富含A/T的DNA寡聚体的相互作用

DOI：
发表时间：
2004
期刊：
J.Biol.Chem. 279(30)
影响因子：
0
作者：
S.Okubo;F.Hara;Y.Tsuchida;S.Shimotakahara;S.Suzuki;H.Hatanaka;S.Yokoyama;H.Tanaka;H.Yasuda;H.Shindo
通讯作者：
H.Shindo

NMR structure of the N-termnal domain of SUMO ligase PLAS1 and its interaction with tumor supressor p53 and A/T-rich DNA oligmers

SUMO 连接酶 PLAS1 N 端结构域的 NMR 结构及其与肿瘤抑制因子 p53 和富含 A/T 的 DNA 寡聚体的相互作用

DOI：
发表时间：
2004
期刊：
J. Biol. Chem 279.30
影响因子：
0
作者：
S.Okubo;F.Hara;Y.Tsuchida;S.Shimotakahara;S.Suzuki;H.hatanaka;S.Yokoyama;H.Tanaka;H.Yasuda;H.Shido
通讯作者：
H.Shido

NMR structure of the N-terminal domain of SUMO ligase PIAS1 and its interaction with tumor suppressor p53 and A/T-rich DNA oligomers