A New E3 Ligase Implicated in Protein Quality Control and Neurodegeneration

一种与蛋白质质量控​​制和神经变性有关的新 E3 连接酶

• 批准号: 8787516
• 负责人: CLAUDIO A.P. JOAZEIRO
• 金额: $ 41.45万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-01-15 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8787516
• 关键词: Afferent Neurons; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animal Model; Basic Science; Binding; Biochemical; Biochemistry; Biological; Biological Assay; Biological Process; Cells; Cellular biology; Defect; Development; Disease; Disease model; Drug Targeting; Eukaryota; Eukaryotic Cell; Exhibits; Gene Expression; Genes; Genetic study; Goals; Homologous Gene; Human; Huntington Disease; In Vitro; Lead; Mammalian Cell; Mammals; Mass Spectrum Analysis; Mediating; Messenger RNA; Modeling; Modification; Molecular; Molecular Chaperones; Molecular Genetics; Motor Neurons; Mus; Mutation; Names; Nerve Degeneration; Neurodegenerative Disorders; Organism; Orthologous Gene; Paralysed; Parkinson Disease; Pathogenesis; Pathway interactions; Poly(A) Tail; Prevalence; Process; Production; Proteins; Public Health; Quality Control; Reaction; Regulation; Reporting; Research; Research Design; Research Proposals; Ribosomes; Role; Saccharomycetales; Signal Transduction; Specific qualifier value; System; Terminator Codon; Testing; Therapeutic Intervention; Tissues; To specify; Translations; Ubiquitin; Ubiquitination; Work; Yeasts; base; early onset; gene discovery; gene function; mRNA Decay; mouse model; novel; novel therapeutics; polypeptide; protein aggregate; protein aggregation; protein degradation; reconstitution; tissue culture; ubiquitin-protein ligase; yeast genetics

7. PROJECT SUMMARY Background and relevance: The goal of this basic research proposal is to better understand the causes of neurodegenerative disorders at the molecular level. Among the best known examples are Alzheimer's, Parkinson's and Huntington's diseases, and Amyotrophic Lateral Sclerosis (ALS). Neurodegenerative diseases are incurable and debilitating conditions with increasing prevalence, and without their pathogenic mechanisms being elucidated in sufficient detail, our ability to generate a rationale for corrective therapeutic interventions is greatly limited. We had previously characterized a novel mouse model of neurodegeneration caused by mutation of the LISTER gene. Consistent with an important biological function, this gene is conserved in all organisms from yeast to humans. Accordingly, we have been utilizing various models to discover the gene function and how defects in this function may lead to the disease. The LISTER gene encodes a protein that acts as an E3 ubiquitin ligase, named Listerin. More recently, using yeast we discovered a function for Listerin that is consistent with a role in neurodegeneration in mammals: it functions in a process known as protein quality control whereby aberrant proteins are targeted for degradation. Specifically, its targets are proteins encoded by defective mRNA lacking stop codons ("non-stop proteins"). Listerin functions by tagging those aberrant proteins with molecules of ubiquitin (ubiquitination), which often acts as a destruction signal. Mutation of Listerin causes those toxic proteins to accumulate. With the proposed research we plan to specify the features of Listerin-mediated ubiquitination and expand the findings towards disease. Objective/Hypothesis: Our main hypotheses are that (a) Listerin acts in protein quality control by ubiquitinating non-stop polypeptides stalled in ribosomes, and (b) defective Listerin-mediated degradation leads to the formation of protein aggregates and inclusions, which are hallmarks of neurodegeneration. The Specific Aims are to characterize Listerin's function in non-stop protein degradation, to investigate the extent of conservation of Listerin's function in protein quality control, and to investigate how defects in this function lead to neurodegeneration. Our long-term objective is to help elucidate molecular mechanisms involved in the pathogenesis of human neurodegenerative disease. Study design: We will use biochemistry and yeast molecular genetics to specify how the E3 recognizes its specific target substrates; mammalian tissue culture and biochemistry to investigate the regulation of non-stop protein degradation by mammalian Listerin and biochemistry and cell biology to study the consequences of non-stop protein accumulation in cells.

7。项目摘要 背景和相关性：这项基础研究建议的目的是更好地了解 分子水平的神经退行性疾病。最著名的例子是阿尔茨海默氏症， 帕金森氏症和亨廷顿疾病，肌萎缩性侧索硬化症（ALS）。神经退行性疾病 无法治愈，并使患病率增加，并且没有致病机制 详细阐明，我们有能力为纠正治疗干预措施产生理由 非常有限。我们以前曾表征由由 李斯特基因的突变。与重要的生物学功能一致，该基因在所有基因中都保存 从酵母到人类的生物。因此，我们一直在利用各种模型来发现基因 功能以及此功能中的缺陷如何导致该疾病。李斯特基因编码一种蛋白质 充当E3泛素连接酶，名为Listerin。最近，使用酵母，我们发现了李斯特蛋白的功能 这与哺乳动物中神经退行性的作用一致：它在称为蛋白质的过程中起作用 质量控制，使异常蛋白的靶向降解。具体而言，其靶标是蛋白质 由缺乏终止密码子（“非停车蛋白”）的缺陷mRNA编码。李斯特蛋白通过标记这些功能 异常的蛋白质具有泛素分子（泛素化），通常充当破坏信号。突变 李斯特蛋白会导致这些有毒蛋白积聚。通过拟议的研究，我们计划指定 李斯特蛋白介导的泛素化的特征并将发现扩大到疾病。 客观/假设：我们的主要假设是（a）李斯特蛋白在蛋白质质量控​​制中通过 在核糖体中停滞不前的泛素化非停车多肽，以及（b）李斯特蛋白介导的缺陷降解 导致蛋白质聚集体和夹杂物的形成，这些蛋白质聚集体是神经变性的标志。 具体目的是表征李斯特蛋白在不间断蛋白质降解中的功能，以研究 李斯特蛋白在蛋白质质量控​​制中的功能的保护程度，并研究了如何缺陷 功能导致神经变性。我们的长期目标是帮助阐明分子机制 参与人神经退行性疾病的发病机理。 研究设计：我们将使用生物化学和酵母分子遗传学来指定E3如何识别其 特定目标基板；哺乳动物组织培养和生物化学研究不间断的调节 哺乳动物李斯特蛋白和生物化学和细胞生物学降解蛋白质降解，以研究的后果 细胞中不间断的蛋白质积累。