Role of Mitophagy in Mitochondrial Protein Quality Control

线粒体自噬在线粒体蛋白质质量控​​制中的作用

• 批准号: 8841573
• 负责人: Adam Lucas Hughes
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-15 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8841573
• 关键词: Acidity; Address; Age; Alzheimer' s Disease; Amino Acids; Autophagocytosis; Award; Biological Assay; Biological Models; Cells; Cellular biology; Chimeric Proteins; Data; Development; Diabetes Mellitus; Disease; Genes; Goals; Health; Human; Individual; Lysosomes; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Methods; Microscopy; Mission; Mitochondria; Mitochondrial Proteins; Molecular; Molecular Target; Nutrient; Organism; Outcome; PINK1 gene; Parkinson Disease; Pathway interactions; Phosphotransferases; Prevention; Process; Proteins; Proteome; Public Health; Quality Control; Recruitment Activity; Research; Role; Saccharomyces cerevisiae; Saccharomycetales; Surface; Techniques; Testing; Time; Translating; United States National Institutes of Health; Vacuole; Work; Yeasts; aged; base; career; coping; early onset; human disease; innovation; insight; loss of function; mitochondrial dysfunction; novel; parkin gene/protein; prevent; programs; protein degradation; research study; response; therapy development; ubiquitin-protein ligase; yeast genetics

PROJECT SUMMARY/ABSTRACT Mitochondrial dysfunction frequently occurs in aged individuals and is a hallmark of diseases such as Parkinson's, Alzheimer's, cancer, and diabetes. Organisms are equipped with mechanisms to cope with mitochondrial dysfunction, including the PINK1-Parkin pathway that promotes lysosomal turnover of mitochondrial proteins by autophagy (mitophagy) in response to mitochondrial dysfunction. However, there is a fundamental gap in understanding how this pathway functions to promote mitochondrial integrity and prevent disease formation. The objective of this application is to determine the mechanism through which damage- induced mitophagy promotes mitochondrial quality control using S. cerevisiae. Previous studies identified a mitophagy pathway in yeast, but this pathway is not homologous to the PINK1-Parkin pathway and does not respond to mitochondrial dysfunction, which limits its usefulness as a model system. The applicant recently discovered a second mitophagy pathway in yeast that is functionally equivalent to the PINK1-Parkin pathway and promotes mitophagy in response to age-induced mitochondrial dysfunction caused by loss of lysosome- like vacuolar acidity. The applicant will use this novel pathway to test the central hypothesis of this application that mitophagy selectively alters the mitochondrial proteome to maintain mitochondrial integrity in response to mitochondrial dysfunction. Understanding how mitophagy prevents mitochondrial dysfunction in yeast will rapidly advance our understanding of this process in humans and facilitate the development of treatments for diseases associated with mitochondrial dysfunction. The objective of this application will be accomplished by pursuing the following three specific aims: Aim 1: Determine how reduced vacuolar acidity causes mitochondrial dysfunction. Suppressor screens and nutrient modulation will be used to characterize pathways that contribute to mitochondrial dysfunction in the absence of vacuolar acidity. Aim 2: Determine how mitochondrial proteins are degraded by mitophagy. This aim will use a microscopy-based mitophagy assay and mass spectrometry to further characterize the damage-induced mitophagy pathway and identify novel genes required for its function. Aim 3: Identify what components of the mitochondria are degraded by mitophagy. This aim will use microscopy-based techniques to identify mitochondrial proteins degraded by mitophagy and determine if mitophagy targets preexisting or newly synthesized proteins. The research proposed in this application is innovative because it dissects the role of mitophagy in mitochondrial quality control using a previously unknown functional equivalent of the PINK1-Parkin pathway in budding yeast and thus brings the power of yeast genetics to the damage-induced mitophagy field. This is significant because it will provide a deep molecular understanding of a mitochondrial protein quality control pathway that protects cells against age-induced mitochondrial dysfunction and prevents the development of Parkinson's disease.

项目摘要/摘要 线粒体功能障碍经常发生在老年人中，是诸如疾病的标志 帕金森氏症，阿尔茨海默氏症，癌症和糖尿病。有机体配备了应对的机制 线粒体功能障碍，包括促进溶酶体周转率的Pink1-Parkin途径 线粒体（线粒体）响应线粒体功能障碍的线粒体蛋白。但是，有一个 理解该途径如何促进线粒体完整性并防止该途径的基本差距 疾病形成。该应用的目的是确定损害的机制 诱导的线粒体使用酿酒酵母促进线粒体质量控制。以前的研究确定了 酵母中的线粒体途径，但该途径与Pink1-Parkin途径不是同源的，也不是 响应线粒体功能障碍，这限制了其作为模型系统的有用性。申请人最近 在酵母中发现了第二个线粒体途径，在功能上等同于Pink1-Parkin途径 并促进线粒体对年龄诱导的线粒体功能障碍的响应 像液泡酸度一样。申请人将使用这种新颖的途径来测试此申请的中心假设 线粒体有选择地改变线粒体蛋白质组，以维持线粒体完整性。 线粒体功能障碍。了解线粒体如何防止酵母中的线粒体功能障碍 迅速提高我们对人类这一过程的理解，并促进了治疗的发展 与线粒体功能障碍相关的疾病。该应用程序的目的将由 追求以下三个特定目的：目标1：确定液泡酸度降低是如何导致的 线粒体功能障碍。抑制剂筛选和营养调制将用于表征途径 在没有液泡酸度的情况下，这会导致线粒体功能障碍。目标2：确定如何 线粒体蛋白被线粒体降解。此目标将使用基于显微镜的线粒体测定法和 质谱法进一步表征损伤引起的线粒体途径并识别新基因 其功能所需。目标3：确定线粒体的哪些组成部分被线粒体降解。这 AIM将使用基于显微镜的技术来识别线粒体蛋白，并通过线粒体和线粒体降解 确定线粒体是否靶向先前存在或新合成的蛋白质。这项研究提出了 应用具有创新性，因为它剖析了线粒体在线粒体质量控制中的作用 以前未知的功能等效于芽酵母中的Pink1-Parkin途径，因此带来了 酵母遗传学对损伤引起的线粒体场的功率。这很重要，因为它将提供 对线粒体蛋白质质量控​​制途径的深层分子理解，该途径可保护细胞免受 年龄引起的线粒体功能障碍并防止帕金森氏病的发展。