Cell Mechanisms of Abnormal Protein Aggregation

蛋白质异常聚集的细胞机制

• 批准号: 7271879
• 负责人: Michael Y Sherman
• 金额: $ 35.41万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7271879
• 关键词: Actins; Affect; Amyotrophic Lateral Sclerosis; Binding; Biochemical Genetics; Caenorhabditis elegans; Cells; Defect; Disease; Electron Microscopy; Endocytosis; Endocytosis Inhibition; Fluorescence Resonance Energy Transfer; Genes; Genetic Screening; Goals; Homologous Gene; Investigation; Longevity; Mammalian Cell; Mediating; Modeling; Mutation; Nature; Neurodegenerative Disorders; Neurons; Numbers; Parkinson Disease; Play; Poly(A)-Binding Protein II; Prions; Process; Proline-Rich Domain; Proteins; Research; Role; SH3 Domains; Site; Testing; Toxic effect; Tube; Work; Yeast Model System; Yeasts; alpha synuclein; base; human Huntingtin protein; human SNCAIP protein; in vivo; mutant; nervous system disorder; neurotoxicity; polyglutamine; polypeptide; programs; protein aggregation; research study

DESCRIPTION (provided by applicant): Neuronal accumulation of various mutant or damaged proteins results in several neurodegenerative disorders, including Parkinson's disease, ALS, and polyglutamine expansion disorders. Toxic abnormal species can aggregate in cells, and there is an ongoing discussion of how protein aggregation influences neurotoxicity. It has recently become clear that in contrast to protein aggregation in a test tube, aggregation of damaged or mutant polypeptides in vivo is a complicated and tightly regulated process that involves many cellular factors. Using a yeast model of polyglutamine (polyQ) expansion disorders, the PI has carried out a number of genetic screens and found that mutations in several components of the machinery that organizes cortical actin patches (CP) dramatically reduce polyQ aggregation. Furthermore, elimination of CP by arp2 or arp3 mutations completely blocks aggregation of polyQ in cells. This proposal will test the hypothesis that cortical actin patches play a direct role in polyQ aggregation. Accordingly, investigations will be carried out to determine if polyQ-containing polypeptides aggregate at CP sites, and the role of Rnq1 prion in these interactions. The role of components of CP and factors responsible for formation of actin cables in polyQ aggregation will be evaluated. An important goal would be to establish whether CP play a general role in protein aggregation, including aggregation of distinct proteins important for neurological disorders, e.g. synphilin 1, alpha-synuclein and PABP2, and in formation of yeast prions. A critical question will be whether homologs of major components of CP play similar role in polyQ aggregation in mammalian cells. A special focus will be to evaluate a hypothesis that interactions between CP and polyQ are mediated by certain SH3-domain proteins, e.g. Sla1, Rvs167, Bem1 or Hof1. Previous work from the PI's lab showed that polyQ aggregation causes early defect in endocytosis in yeast and mammalian cells. In a separate aim we will test a hypothesis that polyQ aggregation causes inhibition of endocytosis in neurons, using a C. elegans model. It will also be established whether mutations that increase the lifespan and delay the onset of polyQ aggregation in worms also delay the onset of endocytosis defects. Exploration of fundamental mechanisms of protein aggregation that we undertake in this project will help to understand the nature of several neurological disorders.

描述（由申请人提供）： 各种突变或受损蛋白质的神经元积累会导致几种神经退行性疾病，包括帕金森氏病，ALS和聚谷氨酰胺扩张障碍。毒性异常物种可以在细胞中汇总，并且存在持续的讨论蛋白质聚集如何影响神经毒性。最近很明显，与试管中的蛋白质聚集相反，体内受损或突变多肽的聚集是一个复杂且严格调节的过程，涉及许多细胞因子。 PI使用聚谷氨酰胺（PolyQ）扩展疾病的酵母模型进行了许多遗传筛选，发现在组织皮质肌动蛋白斑块（CP）的几个组件中的突变大大降低了Polyq聚集。此外，通过ARP2或ARP3突变消除CP完全阻止了Polyq在细胞中的聚集。该提案将检验以下假设：皮质肌动蛋白斑块在PolyQ聚集中起着直接的作用。因此，将进行研究，以确定在CP位点含有PolyQ的多肽聚集体以及RNQ1 Prion在这些相互作用中的作用。将评估CP组件的作用以及负责在PolyQ聚集中形成肌动蛋白电缆的因素。一个重要的目标是确定CP是否在蛋白质聚集中起一般作用，包括对神经系统疾病重要的不同蛋白质的聚集，例如同步蛋白1，α-突触核蛋白和PABP2，以及在酵母菌prions的形成中。一个关键的问题将是CP主要组成部分的同源物是否在哺乳动物细胞中PolyQ聚集中起相似的作用。一个特别的重点是评估一个假设，即CP和PolyQ之间的相互作用是由某些SH3核蛋白蛋白（例如SLA1，RVS167，BEM1或HOF1。 PI实验室的先前工作表明，PolyQ聚集会导致酵母和哺乳动物细胞内吞作用的早期缺陷。在一个单独的目的中，我们将使用秀丽隐杆线虫模型检验一个假设，即PolyQ聚集会导致神经元内内吞作用的抑制。还将确定蠕虫中PolyQ聚集的突变是否也会延迟内吞作用缺陷的发作。我们在该项目中进行的蛋白质聚集的基本机制的探索将有助于了解几种神经系统疾病的性质。