Structural basis for protein functional transfer by SUMOylation

SUMO化蛋白质功能转移的结构基础

• 批准号: 18370040
• 负责人: SHIRAKAWA Masahiro
• 金额: $ 10.9万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18370040/
• 关键词: SUMO; protein modification; structure; NMR; DNA methylation; X線結晶構造解析法; DNA結合たんぱく質

Human Heat Shock Factor 2 (hHSF2) has been known to be SUMOylated within its DNA binding domain and trimerization domain. Both positive and negative effect on DNA binding activity by SUMOylation had been reported. In order to investigate the mechanism of regulation of DNAbinding activity of hHSF2 by SUMOylation, we analyzed DNA binding activity of the DNA binding domain (DBD) of hHSF2 and observed the effect of SUMOylation. The result indicated that SUMOylation decreases DNA binding activity of hHSF2 DBD. The results of NMR and pulse ESR experiments and model building suggest that the SUMO moiety on SUMO-attached hHSF2 DBD is not fixed, and thus diversely positioned relative to hHSF2 DBD, but the SUMO moiety tend to be in proximity to the putative DNA binding surface of hSF2 DBD. We proposed the attached SUMO stochastically inhibits DNA access.We have determined NMR structure of complex of SUMO-3 and the SUMO-interacting motif (SIM) of MCAF1. The structure implies the existence of two interfaces, which mediate mainly hydrophobic and electrostatic interactions, respectively. The latter might be involved in the preferential binding to SUMO-2/3 over SUMO-1 by MCAF1 SIM.We have determined the crystal structure of the SRA domain of UHRF1 in complex with hemi-methylated DNA. In the complex, a methyl base is flipped out, and is inserted into a pocket of SRA. The structure implies that the flip out mechanism is important for methyl base recognition.

已知人热休克因子2(HHSF2)在其DNA结合区和三聚区内发生SUMO化。糖基化对DNA结合活性既有积极的影响，也有消极的影响。为了探讨SUMO化对hHSF2 DNA结合活性的调节机制，我们分析了hHSF2的DNA结合域(DBD)的DNA结合活性，并观察了SUMO化的影响。结果表明，SUMO化降低了hHSF2 DBD的DNA结合活性。核磁共振和脉冲ESR实验和建模的结果表明，连接在相扑上的hHSF2 DBD上的相扑部分不是固定的，因此相对于hHSF2 DBD有不同的定位，但相扑部分倾向于接近hSF2 DBD的DNA结合面。确定了SUMO-3复合体的核磁共振结构和MCAF1的SUMO相互作用基序(SIM)。该结构意味着存在两个界面，分别主要介导疏水和静电相互作用。后者可能参与了MCAF1 SIM优先结合SUMO-2/3而不是SUMO-1的过程。我们测定了uhrf1的SRA结构域与半甲基化DNA的结合。在该复合体中，一个甲基碱基被翻转出来，并被插入到SRA的口袋中。这一结构表明，翻转机制对于甲基碱基的识别是重要的。

项目成果

Magnetic resonance-based visualization of gene expression in mammalian cells using a bacterial polyphosphate kinase reporter gene

• DOI: 10.2144/000112319
• 发表时间: 2007-02-01
• 期刊: BIOTECHNIQUES
• 影响因子: 2.7
• 作者: Ki, Sewon;Sugihara, Fuminori;Kokubo, Tetsuro
• 通讯作者: Kokubo, Tetsuro

Structure of the SUMO-interacting motif of MBD1-containing chromatin associated factor 1 (MCAF1) bound to SUMO-3.

与 SUMO-3 结合的包含 MBD1 的染色质相关因子 1 (MCAF1) 的 SUMO 相互作用基序的结构。

• 发表时间: 2008
• 期刊: Journal of Biological Chemistry 283
• 作者: Sekiyama;N.;Ikegami;T.;Yamane;T.;Ikeguchi;M.;Uchimura;Y.;Baba;D.;Ariyoshi;M.;Tochio;H.;Saitoh;H.;Shirakawa;M.
• 通讯作者: M.

Hemi-methylated DNA recognition by the SRA protein Np95 via a base flipping mechanism.

SRA 蛋白 Np95 通过碱基翻转机制识别半甲基化 DNA。

• 发表时间: 2008
• 期刊: Nature 455
• 作者: Arita;K.;Ariyoshi;M.;Tochio;H.;Nakamura;Y.;Shirakawa;M.
• 通讯作者: M.

Characterization of the sequence specificity of the RlBm endonuclease domain by structural and biochemical studies.

通过结构和生化研究表征 RlBm 核酸内切酶结构域的序列特异性。

• 发表时间: 2007
• 期刊: Nucleic Acid Research 35
• 作者: Maita;N.;Aoyagi;H.;Osanai;M.;Shirakawa;M.;Fujiwara;H.,
• 通讯作者: H.,

SUMOによるタンパク質修飾生化学79

使用 SUMO 79 进行蛋白质修饰生物化学

• 发表时间: 2007
• 作者: 磯貝信;白川昌宏
• 通讯作者: 白川昌宏