Function of the Six ATPases in the Yeast 19S Particle

酵母 19S 颗粒中六种 ATP 酶的功能

• 批准号: 6935767
• 负责人: Gary L. Kleiger
• 金额: $ 4.99万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6935767
• 关键词: adenosine triphosphate; adenosinetriphosphatase; analog; chemical genetics; enzyme activity; enzyme inhibitors; enzyme substrate; fungal genetics; fungal proteins; gene expression; gene mutation; intracellular transport; membrane transport proteins; microorganism culture; postdoctoral investigator; proteasome; protein binding; protein degradation; protein folding; protein structure function; protein transport; site directed mutagenesis; ubiquitin; yeasts

DESCRIPTION (provided by applicant): The elucidation of the ubiquitin/proteasome system in eukaryotes has greatly contributed to our understanding of how cells work. However, many mechanistic features of how 26S proteasomes function remain unclear. One key question involves the function of the six AAA ATPases found in the 19S regulatory particle of the proteasome. Despite the importance of these proteins in proteasome function, very few studies have directly addressed the specific roles of ATPase activity in proteasome function. The goal of this proposal is to apply a novel 'chemical genetics' approach that will enable us to systematically deduce the molecular function of each ATPase in the 19S. Inhibitors are generated by chemically modifying purine derivatives with bulky substituents to complement a rationally designed enlargement of the ATP-binding sites of the target ATPases. While the 'analogue-sensitive' ATPases can bind to either normal ATP or the ATP analogue, wild-type ATPases are unaffected in the presence of ATP analogue, allowing specific inhibition of a given ATPase in both in vitro and in vivo assays. Initially we will characterize the roles of the yeast 19S ATPases Rpt1 and Rpt5 in proteasome function, with the goal of eventually characterizing all six ATPases.

描述（由申请人提供）：真核生物中泛素/蛋白酶体系统的阐明极大地有助于我们对细胞的工作方式的理解。但是，26S蛋白酶组功能的许多机械特征尚不清楚。一个关键问题涉及在蛋白酶体的19S调节粒子中发现的六个AAA ATPases的功能。尽管这些蛋白质在蛋白酶体功能中很重要，但很少有研究直接解决了ATPase活性在蛋白酶体功能中的特定作用。该提案的目的是采用一种新颖的“化学遗传学”方法，该方法将使我们能够系统地推断出19S中每个ATPase的分子功能。抑制剂是通过化学修饰的嘌呤衍生物具有大量取代基的，以补充靶ATPases的ATP结合位点的合理设计的增大。尽管“模拟敏感” ATPases可以与正常ATP或ATP类似物结合，但在ATP模拟存在下，野生型ATPase不受影响，从而可以在体外和体内分析中特定抑制给定ATPase。最初，我们将表征酵母19S ATPASE RPT1和RPT5在蛋白酶体功能中的作用，其目的是最终表征所有六个ATPASE。