Membrane trafficking to lysosomes

膜转运至溶酶体

• 批准号: 10620966
• 负责人: CHARLES G ODORIZZI
• 金额: $ 45.4万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-04 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10620966
• 关键词: Address; Biogenesis; Biological Process; Cell Surface Receptors; Cell physiology; Cells; Collaborations; Diagnostic; Disease; Endocytosis; Endosomes; Enzymes; Foundations; Functional disorder; Genetic; Genetic Diseases; Genetic Screening; HIV-1; Human; In Vitro; Infection; Link; Lumen of the Lysosome; Lysosomes; Mediating; Membrane; Microscopy; Mission; Molecular; Mutation; National Institute of General Medical Sciences; Pathway interactions; Physiological; Prevention; Process; Productivity; Protein Analysis; Proteins; Proteomics; Public Health; Regulation; Research; Resolution; Saccharomyces cerevisiae; Saccharomycetales; Site; Sorting; System; Techniques; Ubiquitin; Vesicle; Vision; Work; Yeasts; biophysical analysis; disease diagnosis; extracellular; man; model organism; novel; pathogen; pathogenic virus; proteostasis; receptor; reconstitution; tool; trafficking; vesicle transport

PROJECT SUMMARY Lysosomes are the primary catabolic site of cells, serving to degrade extracellular material internalized by en- docytosis and intracellular components earmarked for turnover. These items and the hydrolytic enzymes that degrade them are delivered to lysosomes by multiple membrane trafficking pathways, which are operated by cellular protein machineries that are evolutionarily conserved from yeast to man. Mutations that disrupt the ac- tivities of these machineries are linked to genetic diseases, and pathogens exploit these trafficking pathways to establish infection. Many of the molecular mechanisms that operate membrane trafficking machineries are un- known. My lab addresses fundamental questions about membrane trafficking to lysosomes using the budding yeast Saccharomyces cerevisiae as a genetically tractable model organism. The major questions we plan to address over the next five years include the following. 1) What are the physiological mechanisms that regulate the trafficking of cell-surface receptors to the hydrolytic interior of the lysosome? This trafficking pathway func- tions at endosomes to sort endocytosed receptors into membrane-enclosed transport vesicles that are deliv- ered into the lysosome lumen. Recent progress from my lab has revealed that the formation of these vesicles is coordinated with other processes that are vital to cellular physiology, including ubiquitin protein homeostasis and endocytic pH regulation. We plan to identify the machineries that interface these different cellular systems and determine how they exert control over the protein machinery that mediates the formation of vesicles trans- ported to the lysosome lumen. These results will define ways in which receptor degradation is coordinated with cellular physiology. 2) What are the mechanisms that mediate the delivery of newly synthesized transmem- brane proteins that are destined to function at the lysosomal membrane? Using novel genetic tools we created for discovery and diagnostics, we recently identified two specific cellular machineries and several additional candidate machineries that function in this trafficking pathway. We plan to define the mechanisms by which these machineries function in transport, which will establish fundamental principles that underly the biogenesis of lysosomes. Our work is bolstered by ongoing productive collaborations that employ diverse interdisciplinary techniques, including high-resolution microscopy, biophysical analyses of protein assemblies reconstituted in vitro, proteomics, and genetic screening. Moving forward, our overall vision is to continue exploiting yeast for discovering and mechanistically understanding membrane trafficking to lysosomes while also addressing the extent to which these mechanisms are conserved in human cells. The information gained from this work will provide an understanding of how membrane trafficking pathways to lysosomes operate under normal physio- logical conditions and how they are vulnerable in disease states.

项目摘要 溶酶体是细胞的主要分解代谢部位，可降解由en- 致力于营业额的密码和细胞内组件。这些物品和水解酶 降解它们是由多个膜运输途径传递到溶酶体的，这些途径由 从酵母到人的进化保守的细胞蛋白质机制。破坏AC-的突变 这些机械的琐事与遗传疾病有关，病原体利用这些贩运途径 建立感染。许多操作膜运输机制的分子机制都是 已知。我的实验室解决了使用萌芽 酿酒酵母作为一种遗传处理模型生物。我们计划的主要问题 未来五年的地址包括以下内容。 1）调节的生理机制是什么 将细胞表面受体运输到溶酶体的水解内部？这个贩运途径功能 - 内体的tions将内吞受体分类为膜封闭的转运囊泡，这些囊泡 进入溶酶体腔。我实验室的最新进展表明，这些囊泡的形成 与其他对细胞生理至关重要的过程协调，包括泛素蛋白稳态 和内吞pH调节。我们计划识别连接这些不同蜂窝系统的机器 并确定它们如何对蛋白质机制施加介导囊泡形成的蛋白质机制的控制 移植到溶酶体腔。这些结果将定义受体降解与与 细胞生理。 2）介导新合成的传输的传递的机制是什么 注定要在溶酶体膜上起作用的brane蛋白？使用我们创建的新型遗传工具 对于发现和诊断，我们最近确定了两个特定的蜂窝机械和其他几个 在此贩运途径中起作用的候选机器。我们计划定义该机制 这些机器在运输中起作用，该运输将建立基本原理 溶酶体。我们的工作得到了持续的生产合作的支持，这些合作采用多样化的跨学科 技术，包括高分辨率显微镜，对蛋白质组件的生物物理分析。 体外，蛋白质组学和基因筛查。向前迈进，我们的整体愿景是继续利用酵母 发现并机械理解膜运输到溶酶体，同时也解决了 这些机制在人类细胞中保守的程度。从这项工作中获得的信息将 提供了解溶酶体在正常物理学下运作的膜运输途径的理解 逻辑条件及其在疾病状态中的脆弱性。