MECHANISM OF CHAPERONE-ASSISTED ASSEMBLY OF PROTEASOME REGULATORY PARTICLE

分子伴侣辅助蛋白酶体调控颗粒的组装机制

• 批准号: 8359664
• 负责人: Jeroen Roelofs
• 金额: $ 26.46万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8359664
• 关键词: ATP phosphohydrolase; Apoptosis; Binding; Biological Assay; Cell division; Cell physiology; Cells; Complex; DNA Repair; Disease; Drug Delivery Systems; Funding; Genetic Transcription; Goals; Grant; Homeostasis; In Vitro; Molecular Chaperones; Multiple Myeloma; National Center for Research Resources; Neurodegenerative Disorders; Peptide Hydrolases; Play; Principal Investigator; Process; Property; Proteins; Research; Research Infrastructure; Resources; Role; Source; Therapeutic; United States National Institutes of Health; Work; base; cost; multicatalytic endopeptidase complex; particle; prevent; protein degradation; protein structure function; reconstitution; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The proteasome is the major cellular protease. It is involved in the controlled degradation of proteins that regulate a wide variety of cellular processes, such as transcription, apoptosis, cell division and DNA repair. With an important role in homeostasis of so many proteins it is not surprising that observed increased proteasome activity (e.g. in multiple myelomas) or decreased proteasome activity (e.g. in many neurodegenerative diseases) is a pathological factor in many diseases. One determinant of cellular proteasome activity is the level of proteasomes in the cell. Thus, it is important to understand how cellular proteasome levels are regulated and how proteasome assembly is regulated. The long-term goal of this project is to understand the mechanisms of proteasome assembly. Recent work has identified four chaperones that facilitate the formation of proteasome regulatory particle (RP). Each chaperone binds to the C-domain of a specific AAA-ATPases located in the RP. Despite this similarity in function and binding properties, there is no sequence or structural conservation among these chaperones. The objective of the research proposed here is to understand the role each RP chaperone plays in the formation of proteasomes. We hypothesize that the chaperones act as templates for specific base subunits in the assembly of RP. Secondly, they regulate the order in which precursor complexes assemble in a spatial and temporal manner. We also hypothesize that the structural difference among the chaperones are important to accommodate as well as prevent a unique set of interactions for specific ATPases during the assembly process. We will use in vitro binding and reconstitution assays to study this. We furthermore will obtain structural information of the chaperones in combination with the C-domain they bind to. We expect that information from the experiments proposed here will provide a detailed understanding of the mechanisms the chaperones employ to assist proteasome assembly. This could potentially provide new drug targets and ultimately enable us to manipulate proteasome levels or activity for therapeutic means.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 蛋白酶体是主要的细胞蛋白酶。它参与蛋白质的受控降解，这些蛋白质调节多种细胞过程，如转录、凋亡、细胞分裂和DNA修复。由于在如此多的蛋白质的体内平衡中具有重要作用，因此观察到的蛋白酶体活性增加（例如在多发性骨髓瘤中）或蛋白酶体活性降低（例如在许多神经退行性疾病中）是许多疾病中的病理因素并不奇怪。细胞蛋白酶体活性的一个决定因素是细胞中蛋白酶体的水平。因此，重要的是要了解细胞蛋白酶体水平是如何调节的，以及蛋白酶体组装是如何调节的。该项目的长期目标是了解蛋白酶体组装的机制。最近的工作已经确定了四个分子伴侣，促进蛋白酶体调节颗粒（RP）的形成。每个分子伴侣与位于RP中的特定AAA-ATP酶的C结构域结合。尽管在功能和结合特性上有这种相似性，但这些分子伴侣之间没有序列或结构保守性。 本研究的目的是了解每个RP分子伴侣在蛋白酶体形成中的作用。我们假设分子伴侣在RP组装中充当特定碱基亚基的模板。其次，它们调节前体复合物在空间和时间上组装的顺序。我们还假设，分子伴侣之间的结构差异是很重要的，以适应以及防止一组独特的相互作用，为特定的ATP酶在组装过程中。我们将使用体外结合和重建测定来研究这一点。此外，我们将获得分子伴侣的结构信息与它们结合的C结构域的组合。我们希望从这里提出的实验信息将提供一个详细的了解的机制，伴侣协助蛋白酶体组装。这可能提供新的药物靶点，并最终使我们能够操纵蛋白酶体水平或活性用于治疗手段。