Signal transduction by a yeast two-component regulator

酵母双组分调节剂的信号转导

• 批准号: 6437148
• 负责人: JAN S. FASSLER
• 金额: $ 30.28万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-01-01 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6437148
• 关键词: DNA; DNA footprinting; Saccharomyces cerevisiae; aspartate; biological signal transduction; cell osmotic pressure; chimeric proteins; fungal genetics; fungal proteins; gene environment interaction; gene expression; gene mutation; genetic regulation; genetic regulatory element; green fluorescent proteins; histidine; intermolecular interaction; intracellular; mass spectrometry; molecular site; phosphorylation; protein kinase; protein localization; protein structure function; protein transport; transcription factor

DESCRIPTION (Provided by applicant): The osmotic response in the yeast is mediated in part by the HOG1 MAP kinase cascade, a signal transduction pathway whose activity is regulated by members of the two-component signal transduction family. Sln1p is a plasma-membrane localized two-component sensor-kinase that autophosphorylates on a conserved histidine upon stimulation. The Sln1p phosphoryl group is relayed to aspartyl and histidyl groups on sln1p and Ypd1p respectively and finally to aspartyl groups on the two response regulators, Ssk1p and Skn7p. The phosphorylation state of Ssk1p regulates HOG1 pathway activity, while Skn7p is a bifunctional transcription factorwhose phosphorylation state dictates whether osmotic response or oxidative response genes will be activated. Our isolation and characterization of a set of three s1nl* mutants that cause a constitutive kinase phenotype and activation of the Skn7p response regulator have focused our attention on various aspects of the SLN1-YPD1-SKN7 phosphorelay. In this proposal we discuss the molecular basis of Sln1 kinase activation, the details of signal propagation and the molecular basisfor regulation of Skn7p function by phosphorylation. The specific aims of this grant are: (1) to investigate the role of a cytoplasmic coiled-coil region in regulating Sln1 kinase activity using activating sln1* mutations and SLN1 chimeras as tools to dissect the process; (2) to investigate how thesln1p signal is propagated to the nucleus by careful analysis ofSkn7p and Ypd1p localization and by defining the cis-and trans-acting requirements for their localization; and (3) to investigate the molecular details of SLN1-SKN7 dependent transcriptional events by analysis of DNA and protein interaction with constitutive and non-phosphorylatable forms of the Skn7 protein. The power of genetics approaches has made the yeast S. cerevisiae an important model for the study of stress response. Regulation of the yeast osmotic stress pathway by a two-component type histidine kinase introduces added health implications since, given the restricted presence of two-component systems in lower eukaryotes and plants, the histidine kinase are an excellent potential anti-fungal drug target.

描述（由申请人提供）：酵母的渗透响应是 部分由 HOG1 MAP 激酶级联介导，这是一种信号转导途径 其活性受到二元信号转导成员的调节 家庭。 Sln1p 是一种质膜局部双组分传感器激酶， 刺激后在保守的组氨酸上自磷酸化。 Sln1p 磷酰基被转移到 sln1p 和 Ypd1p 上的天冬氨酰基和组氨酰基上 分别最后对两个反应调节剂进行天冬氨酰基团， Ssk1p 和 Skn7p。 Ssk1p的磷酸化状态调节HOG1通路 活性，而 Skn7p 是一种双功能转录因子，其 磷酸化状态决定渗透反应还是氧化反应 基因将被激活。我们对一组三者的分离和表征 s1nl* 突变体导致组成型激酶表型和激活 Skn7p 响应调节器已将我们的注意力集中在 SLN1-YPD1-SKN7磷继电器。在这个提案中，我们讨论了分子基础 Sln1 激酶激活、信号传播的细节和分子机制 通过磷酸化调节 Skn7p 功能的基础。具体目标 该资助的目的是：（1）研究细胞质卷曲螺旋区域的作用 通过激活 sln1* 突变和 SLN1 来调节 Sln1 激酶活性 嵌合体作为剖析该过程的工具； (2) 调查thesln1p如何 通过仔细分析 Skn7p 和 Ypd1p 将信号传播到细胞核 本地化并通过定义其顺式和反式作用要求 本土化; (3) 研究SLN1-SKN7的分子细节 通过 DNA 和蛋白质相互作用分析依赖转录事件 具有组成型和非磷酸化形式的 Skn7 蛋白。电源 遗传学方法的发展使酿酒酵母成为重要的模型 应激反应的研究。酵母渗透胁迫途径的调节 双组分组氨酸激酶引入了额外的健康影响 因为，鉴于低层地区双组分系统的存在受到限制 真核生物和植物中，组氨酸激酶具有极好的潜力 抗真菌药物靶点。