Signal transduction by a yeast two-component regulator

酵母双组分调节剂的信号转导

基本信息

批准号：
6841623
负责人：
JAN S. FASSLER
金额：
$ 30.39万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-01-01 至 2006-12-31
项目状态：
已结题

项目摘要

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）：酵母中的渗透反应是由HOG1 MAP激酶级联反应的部分介导的信号转导途径其活性受两分量信号转导的成员的调节家庭。 SLN1P是一种等离子体膜的局部局部化的两种传感器激酶刺激后，自磷酸化在保守的组氨酸上。 SLN1P 磷酸基在SLN1P和YPD1P上转移到Aspartyl和组酰基中分别和最后是两个响应调节剂的天冬氨酸， SSK1P和SKN7P。 SSK1P的磷酸化状态调节HOG1途径活动，而SKN7P是双功能转录因子磷酸化状态决定渗透反应还是氧化反应基因将被激活。我们隔离和表征三个 S1NL*引起组成型激酶表型的突变体和激活 SKN7P响应调节器将我们的注意力集中在 SLN1-ippd1-SKN7磷层。在此提案中，我们讨论了 SLN1激酶激活，信号传播的细节和分子的细节通过磷酸化对SKN7P功能的基础调节。具体目的该赠款是：（1）调查细胞质卷曲圈区域的作用使用激活SLN1*突变和SLN1调节SLN1激酶活性嵌合体作为解剖过程的工具；（2）研究如何thesln1p 通过仔细分析SKN7P和YPD1P，信号传播到核。本地化并通过定义cis和跨性别要求本土化; （3）研究SLN1-SKN7的分子细节通过分析DNA和蛋白质相互作用的依赖转录事件 SKN7蛋白的组成型和非磷酸化形式。力量遗传学方法使酵母菌酿酒酵母成为重要模型压力反应的研究。调节酵母渗透应激途径两种组分类型的组氨酸激酶引入了添加的健康影响由于，鉴于较低的两个组件系统的存在限制真核生物和植物，组氨酸激酶是一种极好的潜力抗真菌药物靶标。