Equipment Supplement: Ubiquitin-Dependent Protein Regulation and Quality Control of the Lysosomal Membrane

设备补充：泛素依赖性蛋白质调节和溶酶体膜的质量控制

• 批准号: 10387872
• 负责人: Ming Li
• 金额: $ 21万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-13 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10387872
• 关键词: Aging; Alzheimer' s Disease; Autophagocytosis; Biological Models; Cells; Complex; Degradation Pathway; Development; Digestion; Down-Regulation; Endocytosis; Equipment; Functional disorder; Goals; Human; Huntington Disease; Investigation; Laboratories; Light; Lysosomal Storage Diseases; Lysosomes; Mammalian Cell; Membrane; Membrane Proteins; Molecular; Neurodegenerative Disorders; Organelles; Parkinson Disease; Pathway interactions; Play; Proteins; Proteome; Proteomics; Quality Control; Recycling; Regulation; Research; Role; Signal Transduction; Stress; Structure; Ubiquitin; Vacuole; Yeasts; cell growth; detection of nutrient; exhaustion; follow-up; lysosome membrane; recruit; treatment strategy; ubiquitin-protein ligase

Project Summary The lysosome is an essential organelle responsible for the digestion and recycling of materials delivered by endocytosis and autophagy. It also plays important roles in nutrient sensing and control of cell growth by regulating the localization and activity of mTORC1 signaling complex. Because of its importance, lysosome dysfunction leads to ~ 50 types of lysosomal storage diseases (LSDs) and contributes to many aging-related neurodegenerative diseases such as Alzheimer's, Huntington's, and Parkinson's diseases. Despite exhaustive research on how proteins are delivered to lysosomes, how lysosomes regulate their own membrane proteins remains poorly understood. However, studying this question will reveal how cells maintain a healthy lysosome during stresses and aging. Our long-term goal is to understand these fundamental questions using both yeast and mammalian cells as model systems. Recently, we discovered a ubiquitin- and ESCRT- dependent down-regulation pathway for lysosome (vacuole) membrane proteins in yeast. Follow-up investigations in our laboratory led us to hypothesize that the ubiquitin- and ESCRT- dependent degradation pathway is a general conserved mechanism to regulate the lysosome membrane composition from yeast to human. Consistently, recent proteomic studies identified multiple E3 ubiquitin ligases on the human lysosome membrane. Furthermore, the ESCRT machinery was shown to be recruited to the human lysosome membrane. In this proposed research, we plan to expand our initial findings by pursuing three specific aims. Our Aim 1 will investigate how TORC1 regulates the vacuole membrane proteome via the ubiquitin- and ESCRT-dependent pathway in yeast. Our Aim 2 will study how yeast vacuole membrane E3 ligases recognize their membrane substrates at both structure and function level. Our Aim 3 will study how human lysosomes turnover their membrane proteins. Our research will shed light on the development of new treatment strategies for LSDs and lysosome-related neurodegenerative diseases.

项目概要 溶酶体是负责物质消化和回收的重要细胞器 通过内吞作用和自噬传递。它还在营养传感和 通过调节 mTORC1 信号复合物的定位和活性来控制细胞生长。 由于其重要性，溶酶体功能障碍会导致约 50 种类型的溶酶体储存 疾病（LSD）并导致许多与衰老相关的神经退行性疾病，例如 阿尔茨海默病、亨廷顿病和帕金森病。尽管对如何 蛋白质被传递到溶酶体，溶酶体如何调节自身的膜蛋白 仍然知之甚少。然而，研究这个问题将揭示细胞如何维持 压力和衰老过程中的健康溶酶体。 我们的长期目标是使用酵母和 哺乳动物细胞作为模型系统。最近，我们发现了一种泛素-和ESCRT- 酵母中溶酶体（液泡）膜蛋白的依赖性下调途径。 我们实验室的后续调查使我们推测泛素和 ESCRT 依赖性降解途径是一种普遍保守的调节机制 从酵母到人类的溶酶体膜成分。一致地，最近的蛋白质组学 研究在人溶酶体膜上发现了多种 E3 泛素连接酶。此外， 研究表明，ESCRT 机制被招募到人类溶酶体膜上。 在这项拟议的研究中，我们计划通过追求三个具体目标来扩展我们的初步发现。 我们的目标 1 将研究 TORC1 如何通过 酵母中泛素和 ESCRT 依赖性途径。我们的目标 2 将研究酵母液泡如何形成 膜 E3 连接酶在结构和功能水平上识别其膜底物。 我们的目标 3 将研究人类溶酶体如何翻转其膜蛋白。我们的研究 将有助于开发 LSD 和溶酶体相关的新治疗策略 神经退行性疾病。