Regulatory Mechanism of Stress-Responsive Signal Transduction Pathway in Human and Yeast Cells

人和酵母细胞应激反应信号转导途径的调控机制

• 批准号: 14086202
• 负责人: SAITO Haruo
• 金额: $ 135.94万
• 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 on Priority Areas
• 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/en/grant/KAKENHI-PROJECT-14086202/
• 关键词: gene; environment; enzyme; signal transduction; stress; 細胞内シグナル伝達; ストレス応答; MAPキナーゼ; ヒト培養細胞; 酵母; 分子生物学; 細胞生物学

1) Regulatory mechanism of human stress-responsive signal transduction pathwayWe revealed the mechanism of MTK1 activation by the stress-inducible Gadd45 proteins. Gadd45 binds to the N-terminal region of the MTK1, induces an N-C dissociation in MTK1 molecule, and thus facilitates MTK1 dimerization that leads to MTK1 activation by auto-phosphorylation. We showed that TGF-beta activates MTK1 through induction of the expression of Gadd45. We also identified a novel docking interaction between stress-activated MAPKKK (SAP3K) and stress-activated MAPKK (SAP2K). We showed that a C-terminal region in SAP2K, termed the DVD domain, was 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.2) Regulatory mechanism of yeast stress-responsive signal transduction pathway.We identified the mucin-like membrane glycoproteins Hkr1 and Msb2 as putative osmosensors in the Sho1 branch of Hog1 MAPK pathway. We found a specific docking interaction between yeast osmoregulatory Ssk2 SAP3K and Pbs2 SAP2K, and showed that the interaction is essential for specific activation of Pbs2 by Ssk2. We also analyzed the functional roles of the docking interaction between Hog 1 and its activator, the Pbs2 SAP2K, as well as that between Hog1 and its inactivator, the Ptp2 protein tyrosine phosphatase. Finally, we found that the membrane protein Sho1, the cytoplasmic protein Ste50, and the small G-protein Cdc42 serve as adaptor proteins in yeast osmoregulatory Hog1 MAPK pathway.

1)人应激反应信号转导通路的调控机制我们揭示了应激诱导的Gadd45蛋白激活MTK1的机制。Gadd45结合MTK1的n端区域，诱导MTK1分子的N-C解离，从而促进MTK1二聚化，从而通过自磷酸化激活MTK1。我们发现tgf - β通过诱导Gadd45的表达激活MTK1。我们还发现了应力激活的MAPKKK （SAP3K）和应力激活的MAPKK （SAP2K）之间一种新的对接相互作用。我们发现SAP2K中的c端区域，称为DVD结构域，对这种相互作用至关重要，并阐明了它在SAP2K激活中的作用。利用DVD结构域的特异性相互作用，我们开发了一种新的FRET探针来观察单个活细胞中SAP3K的活性。2)酵母应激反应信号转导通路的调控机制。我们发现黏液样膜糖蛋白Hkr1和Msb2可能是Hog1 MAPK通路Sho1分支的渗透传感器。我们发现了酵母渗透调节Ssk2 SAP3K和Pbs2 SAP2K之间的特异性对接相互作用，并表明这种相互作用是Ssk2特异性激活Pbs2所必需的。我们还分析了Hog1与其激活因子Pbs2 SAP2K以及Hog1与其失活因子Ptp2蛋白酪氨酸磷酸酶之间对接相互作用的功能作用。最后，我们发现膜蛋白Sho1、细胞质蛋白Ste50和小g蛋白Cdc42是酵母渗透调节Hog1 MAPK通路的衔接蛋白。

项目成果

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

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

Smad-dependent GADD45b expression mediates delayed activation of p38 MAP kinase by TGF-b.

Smad 依赖性 GADD45b 表达介导 TGF-b 对 p38 MAP 激酶的延迟激活。

• 发表时间: 2002
• 期刊: EMBO J. 21
• 作者: Takekawa;M.;Tatebayashi;K.;Itoh;F.;Adachi;M.;Imai;K.;^*Saito;H.
• 通讯作者: H.

The Slnl-Ypdl-Sskl multistep phosphorelay system that regulates an osmosensing MAP kinase cascade in yeast.

Slnl-Ypdl-Sskl 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