Folding and Activation of Client Proteins by the Hsp90 Molecular Chaperone Machine

Hsp90 分子伴侣机器对客户蛋白的折叠和激活

• 批准号: 0744522
• 负责人: Jill Johnson
• 金额: $ 68.14万
• 依托单位: Regents of the University of Idaho
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-15 至 2014-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0744522&HistoricalAwards=false
• 关键词: Folding; Activation; Client; Proteins; Hsp90

Molecular chaperones regulate the folding of cellular proteins from synthesis until they are properly folded or degraded. The majority of molecular chaperones assist the folding of proteins that are mostly unfolded. The abundant, essential molecular chaperone Hsp90 (heat shock protein, 90 kDa) is unique among chaperones because it preferentially interacts with and is required for the activity of proteins that are almost completely folded. The client proteins that require Hsp90 for function have little or no sequence or structural homology, and little is known about how Hsp90 promotes activation of a select group of diverse cellular client proteins. The goal of this project is to identify the sites of interaction between Hsp90 and two very different client proteins. A panel of mutant forms of Hsp90 will be used as tools to identify and describe critical features of Hsp90 required for interaction with a protein kinase and a transcription factor. Once mutations that disrupt the interaction with two distinct client proteins in vivo are identified, further research will determine whether the mutations disrupt the in vitro assembly of Hsp90-client protein complexes using cell lysates and purified components. Additional research will focus on establishing whether select Hsp90 mutations indirectly disrupt client protein interaction by preventing interaction of Hsp90 with regulatory co-chaperone proteins. This research project will lead to clarification of the contact sites between Hsp90 and client proteins and a greater understanding of how Hsp90 and co-chaperones cooperate to promote activation of a diverse group of client proteins. This project will have broad impacts on understanding how Hsp90 interacts with diverse client proteins. This project will enable one graduate student and two undergraduate students per year to conduct independent research projects. All students will have the opportunity to develop skills in organizing and presenting data during laboratory meetings, undergraduate research poster forums and/or additional scientific conferences. Additional graduate and undergraduate students outside of this lab will be exposed to this work through scientific exchanges and seminars among faculty members within the Microbiology, Molecular Biology and Biochemistry department, and the Center for Reproductive Biology, which is a joint program with Washington State University. Through the NIH-INBRE program and the UDOC program that is part of the multi-state WWAMI medical school program, additional undergraduate students and high school students from underrepresented and minority groups will also be exposed to research in the laboratory.

分子伴侣调节细胞蛋白质从合成到正确折叠或降解的折叠。大多数分子伴侣有助于蛋白质的折叠，这些蛋白质大部分是未折叠的。丰富的必需分子伴侣Hsp 90（热休克蛋白，90 kDa）在伴侣中是独特的，因为它优先与几乎完全折叠的蛋白质相互作用并且是其活性所需的。需要热休克蛋白90的客户蛋白的功能有很少或没有序列或结构同源性，并很少知道如何热休克蛋白90促进激活的一组选择不同的细胞客户蛋白。该项目的目标是确定热休克蛋白90和两个非常不同的客户蛋白之间的相互作用的网站。一组突变形式的热休克蛋白90将被用来作为工具，以确定和描述的关键功能的热休克蛋白90需要与蛋白激酶和转录因子的相互作用。一旦鉴定出破坏与体内两种不同客户蛋白相互作用的突变，进一步的研究将确定突变是否破坏使用细胞裂解物和纯化组分的Hsp 90-客户蛋白复合物的体外组装。其他研究将集中在确定是否选择热休克蛋白90突变间接破坏客户蛋白的相互作用，防止热休克蛋白90与监管辅伴侣蛋白的相互作用。该研究项目将导致Hsp 90和客户蛋白之间的接触位点的澄清，以及对Hsp 90和辅助分子伴侣如何合作以促进客户蛋白的不同群体的激活的更深入的理解。该项目将对理解Hsp 90如何与不同的客户蛋白相互作用产生广泛的影响。该项目将使一名研究生和两名本科生每年进行独立的研究项目。所有学生都将有机会在实验室会议，本科生研究海报论坛和/或其他科学会议期间组织和展示数据。本实验室以外的其他研究生和本科生将通过微生物学、分子生物学和生物化学系以及生殖生物学中心（与华盛顿州立大学联合开展的项目）的教员之间的科学交流和研讨会接触这项工作。通过NIH-INBRE计划和UDOC计划，这是多州WWAMI医学院计划的一部分，来自代表性不足和少数群体的其他本科生和高中生也将接触到实验室的研究。