Identifying and characterizing protein quality control mechanisms in the nucleus

识别和表征细胞核中的蛋白质质量控​​制机制

• 批准号: 7661148
• 负责人: Richard George Gardner
• 金额: $ 30.47万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-15 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7661148
• 关键词: Address; Age; Algorithms; Alzheimer' s Disease; Amino Acids; Attention; Binding; Biochemical; Biological Assay; Biology; Cell Nucleus; Cell physiology; Cells; Cerebellar Ataxia; Cytoplasm; Degradation Pathway; Disabled Persons; Disease; Employee Strikes; Genetic; Goals; Grant; Homologous Gene; Human; Huntington Disease; Hybrids; Hydrophobicity; Knowledge; Learning; Lesion; Link; Maps; Mediating; Methods; Mitochondria; Molecular Chaperones; Mutagenesis; Mutation; Names; Neuromuscular Diseases; Nuclear; Nuclear Protein; Nuclear Proteins; Oculopharyngeal Muscular Dystrophy; Operating System; Organism; Parkinson Disease; Pathogenesis; Pathology; Pathway interactions; Peptide Library; Peptides; Phase; Physiology; Positioning Attribute; Process; Proteins; Quality Control; Randomized; Reporter; Role; Saccharomyces cerevisiae; Solid; Spinal; Stretching; Structure; Substrate Interaction; System; Toxic effect; Ubiquitin-Protein Ligase Complexes; Ubiquitination; Work; Yeasts; age related; flexibility; genetic selection; grasp; multicatalytic endopeptidase complex; mutant; protein function; protein misfolding; public health relevance; repaired; ubiquitin-protein ligase

DESCRIPTION (provided by applicant): The accumulation of damaged or misfolded proteins can have devastating effects on cellular physiology and viability. To protect itself, the cell possesses numerous Protein Quality Control (PQC) systems that minimize the persistence and detrimental effects of these aberrant proteins. While a variety of PQC systems have been characterized in the cytoplasm, ER, and mitochondria, how the nucleus manages aberrant proteins is poorly understood. But, understanding this fundamental aspect of nuclear biology is important because many age-correlated neuromuscular disorders (Huntington's, Kennedy's, several spinal-cerebellar ataxias) result from toxic accumulation of aberrant proteins in the nucleus. Therefore, our long-term goals are to understand how the nucleus normally protects itself from toxic aberrant proteins by identifying PQC systems that operate in the nucleus, characterizing how these systems recognize and target aberrant proteins, examining if lesions in nuclear PQC systems are required for these pathologies to develop, and determining the specific nuclear processes that are disrupted by aberrant nuclear proteins to cause toxicity. From our initial studies in S. cerevisiae, we've identified the first-known PQC degradation system that acts in the nucleus. The central player of this nuclear PQC system is San1, a nuclear-localized ubiquitin- protein ligase that targets aberrant nuclear proteins for ubiquitination and proteasome degradation. With the discovery of San1, we're now well positioned to explore how a nuclear PQC degradation system recognizes its substrates, and what it recognizes as aberrant within those substrates. The specific aims for this grant are to: (1) Determine the regions within San1 responsible for substrate targeting. Mutagenesis of San1 will be used to identify in cis regions important for substrate recognition. (2) Determine the features of an aberrant protein recognized by San1. Mutagenesis of San1 substrates will be used to identify regions required for targeting by San1. (3) Identify & characterize additional components of the San1 pathway. A combination of genetic and biochemical approaches will be used to identify additional San1 pathway components. PUBLIC HEALTH RELEVANCE: Many age-correlated neuromuscular disorders (Huntington's, Kennedy's, several spinal-cerebellar ataxias, and oculopharyngeal muscular dystrophy to name a few) result from toxic accumulation of aggregation-prone proteins in the cell's nucleus. Our goals are to understand how the nucleus normally protects itself from toxic aggregation-prone proteins, and why it fails to do so in these and other diseases.

描述（由申请人提供）：受损或错误折叠蛋白质的积累可能对细胞生理学和活力产生破坏性影响。为了保护自己，细胞拥有许多蛋白质质量控制（PQC）系统，以最大限度地减少这些异常蛋白质的持久性和有害影响。虽然已经在细胞质、ER和线粒体中表征了各种PQC系统，但对细胞核如何管理异常蛋白质知之甚少。但是，理解核生物学的这个基本方面是重要的，因为许多与年龄相关的神经肌肉疾病（亨廷顿氏病、肯尼迪氏病、几种脊髓-小脑共济失调）是由细胞核中异常蛋白质的毒性积累引起的。因此，我们的长期目标是通过识别在细胞核中运行的PQC系统，表征这些系统如何识别和靶向异常蛋白质，检查核PQC系统中的病变是否是这些病理发展所需的，并确定异常核蛋白质破坏的特定核过程来了解细胞核通常如何保护自己免受毒性异常蛋白质的影响。从我们在S.在酿酒酵母中，我们已经确定了第一个已知的在细胞核中起作用的PQC降解系统。该核PQC系统的中心参与者是San 1，一种核定位的泛素-蛋白连接酶，其靶向异常核蛋白进行泛素化和蛋白酶体降解。随着San 1的发现，我们现在可以很好地探索核PQC降解系统如何识别其底物，以及它在这些底物中识别出什么异常。该补助金的具体目标是：（1）确定San 1内负责底物靶向的区域。San 1的诱变将用于鉴定对底物识别重要的顺式区域。(2)确定San 1识别的异常蛋白质的特征。San 1底物的诱变将用于鉴定San 1靶向所需的区域。(3)识别和表征San 1途径的其他组分。将使用遗传和生物化学方法的组合来鉴定其他San 1途径组分。公共卫生相关性：许多与年龄相关的神经肌肉疾病（亨廷顿氏病、肯尼迪氏病、几种脊髓-小脑共济失调和眼咽肌营养不良症等）是由细胞核中易聚集蛋白的毒性积累引起的。我们的目标是了解细胞核通常如何保护自己免受有毒的易聚集蛋白的伤害，以及为什么它在这些和其他疾病中不能这样做。