Regulatory mechanisms of the environmental stress-responsive MAP kinase signal pathway

环境应激响应MAP激酶信号通路的调控机制

• 批准号: 14104021
• 负责人: SAITO Haruo
• 金额: $ 72.55万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (S)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14104021/
• 关键词: intracellular signal transduction; stress response; MAP kinase; human cultured cells; budding yeast; 分子生物学; 細胞生物学

1) Functional roles and regulation of human stress-activated MAP kinase signal transduction pathway.We identified a novel docking interaction between stress-activated MAPKKK (SAP3K) and stress-activated MAPKK (SAP2K). We also found that a C-terminal region in SAP2K, termed the DVD domain, is essential for this interaction, and elucidated its role in SAP2K activation. Using the specific interaction through DVD domain, we developed a new FRET probe to visualize SAP3K activity in single living cells. Furthermore, we revealed the mechanism of MTK1 activation by the stress-inducible Gadd45 proteins. Gadd45 binds to the N-terminal region of MTK1 SAP3K, induces an N-C dissociation in. MTK1 molecule, and thus facilitates MTK1 dimerization that leads to MTK1 activation by auto-phosphorylation.2) Functional roles and regulation of yeast stress-activated MAP kinase signal transduction pathway.We identified a specific docking interaction between yeast osmoregulatory Ssk2 SAP3K and Pbs2 SAP2K, and revealed that the interaction is essential for specific activation of Pbs2 by Ssk2. We found that the membrane protein Sho1, the cytoplasmic protein Ste50, and the small G-protein Cdc42 bind, serve as adaptor proteins in yeast osmoregulatory Hog1 MAPK pathway. We also revealed that the mucin-like membrane glycoproteins Hkr1 and Msb2 are putative osmosensors in the Sho1 branch of Hog1 MAPK pathway. We proposed that external osmostress causes a conformation change in the mucin domains of Hkrl/Msb2, and thus induces an interaction between Sho1 and Hkrl/Msb2 to activate the Hog1 MAPK pathway.

1)人类应激激活的MAP激酶信号转导通路的功能和调控我们发现了一种新的应激激活的MAPKKK（SAP 3 K）和应激激活的MAPKK（SAP 2K）之间的对接相互作用。我们还发现，在SAP 2K的C-末端区域，称为DVD结构域，是必不可少的这种相互作用，并阐明其在SAP 2K激活的作用。利用通过DVD结构域的特异性相互作用，我们开发了一种新的FRET探针来可视化单个活细胞中的SAP 3 K活性。此外，我们揭示了应激诱导的Gadd 45蛋白激活MTK 1的机制。Gadd 45与MTK 1的SAP 3 K的N端结合，诱导SAP 3 K的N-C解离。2）酵母应激激活的MAP激酶信号转导通路的功能及调控：我们鉴定了酵母应激激活的MAP激酶信号转导通路中的Ssk 2 SAP 3 K和Pbs 2 SAP 2K之间的特异性对接作用，揭示了这种对接作用是Ssk 2特异性激活Pbs 2的关键。我们发现，膜蛋白Sho 1，细胞质蛋白Ste 50，和小G蛋白Cdc 42结合，作为接头蛋白在酵母菌调节Hog 1 MAPK途径。我们还发现，粘蛋白样膜糖蛋白Hkr 1和Msb 2是Hog 1 MAPK通路Sho 1分支中假定的信号转导传感器。我们提出，外部渗透胁迫导致Hkrl/Msb 2粘蛋白结构域的构象变化，从而诱导Sho 1和Hkrl/Msb 2之间的相互作用，从而激活Hog 1 MAPK途径。

项目成果

Activation of MTK1/MEKK4 by GADD45 through induced N-C dissociation and dimerization-mediated trans autophosphorylaion of the MTK1 kinase domain.

GADD45 通过诱导 N-C 解离和二聚化介导的 MTK1 激酶结构域反式自磷酸化来激活 MTK1/MEKK4。

• 发表时间: 2007
• 期刊: Molecular and Cellular Biology 27
• 作者: Miyake;Zenshi
• 通讯作者: Zenshi

The Sln1-Ypol-Ssk1 multistep phosphorelay system that regulates an osmosensing MAP kinase cascade in yeast

Sln1-Ypol-Ssk1 多步磷酸中继系统调节酵母中的渗透感应 MAP 激酶级联

• 发表时间: 2003
• 期刊: Histidine kinases in signal transduction, edited by Inouye, M., and Dutta, R., Academic Press, San Diego
• 作者: Saito;Haruo
• 通讯作者: Haruo

Transmembrane mucins Hkr1 and Msb2 are putative osmosensors in the SHO1 branch of yeast HOG pathway

• DOI: 10.1038/sj.emboj.7601796
• 发表时间: 2007-08-08
• 期刊: EMBO JOURNAL
• 影响因子: 11.4
• 作者: Tatebayashi, Kazuo;Tanaka, Keiichiro;Saito, Haruo
• 通讯作者: Saito, Haruo

A docking site determining specificity of Pbs2 MAPKK for Ssk2/Ssk22 MAPKKKs in the yeast HOG pathway

• DOI: 10.1093/emboj/cdg353
• 发表时间: 2003-07-15
• 期刊: EMBO JOURNAL
• 影响因子: 11.4
• 作者: Tatebayashi, K;Takekawa, M;Saito, H
• 通讯作者: Saito, H

Structural basis for the function and regulation of the receptor protein tyrosine phosphatase CD45.

• DOI: 10.1084/jem.20041890
• 发表时间: 2005-02-07
• 期刊: JOURNAL OF EXPERIMENTAL MEDICINE
• 影响因子: 15.3
• 作者: Nam, HJ;Poy, F;Saito, H;Frederick, CA
• 通讯作者: Frederick, CA