Hsp40 and Stress Protection

Hsp40 和压力保护

• 批准号: 6744186
• 负责人: DOUGLAS M CYR
• 金额: $ 25.37万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6744186
• 关键词: Saccharomyces cerevisiae; binding sites; calorimetry; conformation; cytoprotection; enzyme linked immunosorbent assay; heat shock proteins; molecular chaperones; nuclear magnetic resonance spectroscopy; physiologic stressor; polymerase chain reaction; protein biosynthesis; protein denaturation; protein folding; protein metabolism; protein protein interaction

DESCRIPTION (provided by applicant): The long-term objective of this project is to utilize the yeast S. cerevisiae as a model system to determine the mechanism by which the cytosolic Hsp40s Sis1 and Ydjl act with Hsp70 Ssal (Ssal) to protect cells from denaturing physiological stresses such as heat-shock. The hallmark of the heat-shock response is the rapid induction of chaperone protein expression and the global suppression of protein synthesis. These events occur to prevent the accumulation of non-native proteins and suppress protein aggregation. Hsp40s function to suppress protein aggregation by binding non-native proteins and delivering them to the polypeptide binding site of HspT0. In aim 1, we propose a series of experiments that are designed to determine the mechanism for substrate binding by Hsp40s. In aim 2, we will investigate how denatured proteins that are bound to Hsp40 are delivered to Hsp70 to facilitate protein refolding. An additional feature of Hsp40 family members is that they have evolved to have different domain structures and can target Hsp70 to catalyze different cellular reactions. We have demonstrated that the Ydjl and Sis1 function in the cytosol to direct Ssal to facilitate different aspects of protein metabolism. In Aim 3 and Aim 4 we propose a series of domain swap experiments between Ydjl and Sis1 that are designed to define the rules by which Hsp70s cellular functions are specified.The outcome of these studies will define basic mechanisms by which Hsp40s function with Hsp70 to protect cells from physiological stress.

描述（由申请人提供）： 该项目的长期目标是利用酿酒酵母作为模型系统，以确定胞质 Hsp40s Sis1 和 Ydjl 与 Hsp70 Ssal (Ssal) 相互作用的机制，以保护细胞免受变性生理应激（例如热）的影响。震惊。热休克反应的标志是伴侣蛋白表达的快速诱导和蛋白质合成的全面抑制。这些事件的发生是为了防止非天然蛋白质的积累并抑制蛋白质聚集。 Hsp40 通过结合非天然蛋白质并将其递送至 HspT0 的多肽结合位点来抑制蛋白质聚集。在目标 1 中，我们提出了一系列实验，旨在确定 Hsp40 与底物结合的机制。在目标 2 中，我们将研究与 Hsp40 结合的变性蛋白质如何传递至 Hsp70 以促进蛋白质重折叠。 Hsp40家族成员的另一个特点是它们已经进化出不同的结构域结构，并且可以靶向Hsp70来催化不同的细胞反应。我们已经证明 Ydjl 和 Sis1 在细胞质中发挥作用，指导 Ssal 促进蛋白质代谢的不同方面。在目标 3 和目标 4 中，我们提出了 Ydjl 和 Sis1 之间的一系列结构域交换实验，旨在定义指定 Hsp70s 细胞功能的规则。这些研究的结果将定义 Hsp40s 与 Hsp70 共同保护的基本机制细胞免受生理压力。