Role of Ca2+/Calcineurin Signaling in S.Cerevisiae

Ca2/钙调磷酸酶信号在酿酒酵母中的作用

基本信息

批准号：
7930987
负责人：
Martha S. Cyert
金额：
$ 11.31万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2010-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7930987
关键词：
Address Affinity Alleles Animal Model Applied Genetics Biochemical Biological Assay Blood Vessels Brain Calcineurin Calcineurin inhibitor Calmodulin Catalytic Domain Cell Wall Cell physiology Cell surface Cells Cyclosporine Development Docking Event FK506 Family Funding Gene Expression Regulation Genetic Genomics Growth Health Heart Heart Hypertrophy Human Hybrids Immune Integral Membrane Protein Learning Mammals Mass Spectrum Analysis Mediating Membrane Memory Methods Mutation Myocardium Organism Point Mutation Property Protein Microchips Protein phosphatase Proteins Proteomics Regulation Research Research Proposals Role Saccharomyces cerevisiae Screening procedure Signal Pathway Signal Transduction Site Stress Substrate Interaction Testing Ubiquitin Variant Yeasts dosage in vivo mutant novel protein complex research study response scaffold success transcription factor

Address Affinity Alleles Animal Model Applied Genetics Biochemical Biological Assay Blood Vessels Brain Calcineurin Calcineurin inhibitor Calmodulin Catalytic Domain Cell Wall Cell physiology Cell surface Cells Cyclosporine Development Docking Event FK506 Family Funding Gene Expression Regulation Genetic Genomics Growth Health Heart Heart Hypertrophy Human Hybrids Immune Integral Membrane Protein Learning Mammals Mass Spectrum Analysis Mediating Membrane Memory Methods Mutation Myocardium Organism Point Mutation Property Protein Microchips Protein phosphatase Proteins Proteomics Regulation Research Research Proposals Role Saccharomyces cerevisiae Screening procedure Signal Pathway Signal Transduction Site Stress Substrate Interaction Testing Ubiquitin Variant Yeasts dosage in vivo mutant novel protein complex research study response scaffold success transcription factor

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项目摘要

DESCRIPTION (provided by applicant): In mammals, calcineurin, the Ca2+/calmodulin dependent protein phosphatase, regulates immune cell activity, promotes heart and blood vessel development, mediates cardiac muscle response to stress, and modulates learning and memory in the brain. Calcineurin inhibitors, FK506 and cyclosporin A, are used clinically as immunosupressants, and, in animal models, reduce cardiac hypertrophy. As a key effector of Ca2+/calmodulin dependent signaling, detailed analysis of calcineurin function has the potential to impact many aspects of human health and development. In S. cerevisiae, calcineurin promotes survival during environmental stress, and in response to cell wall damage. A major role of calcineurin is to dephosphorylate and activate the Crz1p transcription factor, using mechanism analogous to calcineurin regulation of the mammalian transcription factor, NFAT. Additional calcineurin-mediated events that promote yeast survival during environmental stress are less well characterized, and are the focus of this application. This research aims to identify comprehensively the functions and components of calcineurin-dependent signaling pathways. We address these questions using S. cerevisiae, because of many experimental advantages offered by this simple eukaryotic organism, but strive to establish general principles that apply to calcineurin-dependent signaling in all cells. Previously, we exploited a particular feature of calcineurin signaling, i.e. the requirement for calcineurin to interact directly with its substrates via a docking site that is distinct from residues that are dephosphorylated, to identify several new components of calcineurin-mediated signaling pathways including 3 novel substrates: Slm1p, Slm2p and Hph1p. Here we propose to characterize further the mechanism by which calcineurin interacts with its substrates, which is evolutionary conserved. We will also apply genetic, genomic, and proteomic approaches to the identification of additional substrates and regulators of calcineurin. Specifically we will 1) Characterize calcineurin-docking sites in substrates and examine the impact of calcineurin-substrate affinity on signaling. 2) Examine the effect of specific point mutations in calcineurin on substrate interaction. 3) Identify the role of calcineurin interacting proteins in calcineurin signaling pathways. 4) Identify additional calcineurin substrates using novel proteomic and genomic screening methods.

描述（由申请人提供）：在哺乳动物中，钙调蛋白，Ca2+/钙调蛋白依赖性蛋白磷酸酶，调节免疫细胞活性，促进心脏和血管发育，介导心肌对压力的反应，并调节大脑中的学习和记忆。钙调蛋白抑制剂FK506和环孢菌素A在临床上用作免疫启动剂，在动物模型中，可以减少心脏肥大。作为CA2+/钙调蛋白依赖性信号传导的关键效应因子，对钙调神经酶功能的详细分析具有影响人类健康和发育的许多方面的潜力。在酿酒酵母中，钙调神经蛋白会在环境应激期间促进生存，并响应细胞壁损伤。钙调蛋白的主要作用是使用类似于哺乳动物转录因子NFAT的钙调蛋白调节的机制去磷酸化和激活CRZ1P转录因子。在环境压力期间促进酵母生存的其他钙调蛋白介导的事件的表征较低，并且是该应用的重点。这项研究旨在全面识别钙调蛋白依赖性信号通路的功能和组成部分。我们使用酿酒酵母解决了这些问题，因为这种简单的真核生物提供了许多实验优势，但是努力建立适用于所有细胞中钙调蛋白依赖性信号的一般原理。以前，我们利用了钙调神经蛋白信号传导的特定特征，即钙调神经酶通过与脱磷酸化的残基不同的对接位点直接与其底物相互作用，以识别钙调蛋白介导的几个新成分，包括3个新型信号途径，包括3个新颖的底物：SLM1P，SLM1P，SLM1P，SLM2P和HPHM2P和HPH1P和HPH1P和HPH1P和HPH1P和HPH1P和HPH1P和HPH1P和HPH1P。在这里，我们建议进一步表征钙调蛋白与其底物相互作用的机制，即进化保守。我们还将应用遗传，基因组和蛋白质组学方法来鉴定钙调蛋白的其他底物和调节剂。具体而言，我们将1）表征底物中的钙调神经毒素借入位点，并检查钙调神经蛋白 - 基底亲和力对信号传导的影响。 2）检查钙调蛋白中特定点突变对底物相互作用的影响。 3）确定钙调蛋白相互作用蛋白在钙调神经蛋白信号通路中的作用。 4）使用新型的蛋白质组学和基因组筛选方法鉴定其他钙调蛋白底物。