Molecular mechanisms that regulate vesicle formation and transport

调节囊泡形成和运输的分子机制

基本信息

批准号：
10163556
负责人：
Anjon Audhya
金额：
$ 25万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10163556
关键词：
Animals Biochemistry Biogenesis Biological Models Caenorhabditis elegans Cell Differentiation process Cell Proliferation Cells Cellular biology Complex Cytoplasm Defect Diabetes Mellitus Diffusion Disease Endoplasmic Reticulum Endosomes Estrogen receptor positive Eukaryotic Cell Future Genes Genetic Goals Golgi Apparatus Homeostasis Image Immune System Diseases Intervention Intracellular Membranes Laboratories Lead Link Lipids Location Lysosomes Malignant Neoplasms Mediating Membrane Methodology Molecular Movement Mutation Nerve Degeneration Neurodegenerative Disorders Organelles Pathway interactions Play Process Protein Sortings Proteins Receptor Signaling Regulation Research Resolution Sorting - Cell Movement System Transmembrane Transport Tumor Suppression Vesicle Work insight intracellular protein transport protein degradation protein transport structural biology therapeutic target trafficking

项目摘要

PROJECT SUMMARY How the intracellular membrane system of eukaryotic cells is configured and maintained is a fundamental problem in cell biology. Deficiencies in this organization often lead to disease. The overarching goal of my laboratory is to define the molecular mechanisms that regulate membrane dynamics, including vesicle biogenesis, organelle trafficking, and protein sorting in the early secretory and endocytic pathways of metazoan cells. We aim to determine how normal membrane transport contributes to cellular homeostasis and understand the molecular basis for disease states that emerge when trafficking pathways are disrupted. Using a combination of model systems, structural biology, biochemistry, genetics, and high-resolution subcellular imaging, our studies focus on two essential trafficking pathways necessary for protein and lipid export from the endoplasmic reticulum (ER) and protein turnover within lysosomes via a multivesicular endosome (MVE) intermediate. Mutations in several factors that regulate these processes have been implicated in cancer, diabetes, immune dysfunction, and neurodegeneration. Thus, deciphering the fundamental principles underlying the regulation of ER export and MVE biogenesis should facilitate the future identification of therapeutic targets for disease intervention. One major focus of my research has been elucidating how the early secretory pathway is organized. Our findings revealed the existence of a conserved membrane interface, which links subdomains on the endoplasmic reticulum (ER) that produce COPII-coated transport carriers to juxtaposed ER-Golgi intermediate compartments (ERGIC). We identified Trk-fused gene (TFG) as a key constituent of this interface and have shown that its inhibition uncouples ER and ERGIC membranes and leads to the isotropic diffusion of COPII-coated carriers, reducing the efficiency of cargo secretion. My second research focus is aimed at understanding the mechanisms that direct the formation of MVEs, which bud intralumenal vesicles (ILVs) into their interior to sequester membrane-associated cargoes from the cytoplasm. Eventual fusion of MVEs with lysosomes results in the degradation of ILVs and their associated proteins, which plays a key role in tumor suppression by governing the capture and sequestration of signaling receptors. Using C. elegans, we have developed a new, highly simplified, and genetically tractable system to investigate how components of the endosomal sorting complex required for transport (ESCRT) machinery enables the movement of endocytosed cargo to MVEs and ILVs in the context of an intact, developing animal. Our future work will continue to capitalize on evolving methodologies to further establish how COPII-mediated transport and ESCRT-mediated MVE biogenesis are properly regulated. A better understanding of these processes will yield key insights into the homeostatic controls that sustain normal protein trafficking in the secretory and endocytic pathways during cell proliferation and differentiation.

项目概要真核细胞的细胞内膜系统如何配置和维持是一个基础细胞生物学问题。该组织的缺陷常常导致疾病。我的总体目标实验室的目标是定义调节膜动力学的分子机制，包括囊泡后生动物早期分泌和内吞途径中的生物发生、细胞器运输和蛋白质分选细胞。我们的目标是确定正常的膜运输如何促进细胞稳态并了解当贩运途径被破坏时出现的疾病状态的分子基础。使用组合我们的研究涉及模型系统、结构生物学、生物化学、遗传学和高分辨率亚细胞成像重点关注内质网蛋白质和脂质输出所需的两条重要运输途径 (ER) 和溶酶体内的蛋白质通过多囊泡内体 (MVE) 中间体进行周转。突变在调节这些过程的几个因素与癌症、糖尿病、免疫功能障碍、和神经退行性变。因此，解读 ER 出口监管的基本原则 MVE 的生物发生应有助于未来确定疾病干预的治疗靶点。一我研究的主要重点是阐明早期分泌途径是如何组织的。我们的发现揭示了保守膜界面的存在，该界面连接内质上的子结构域网状结构 (ER)，产生 COPII 涂层的运输载体到并置的 ER-高尔基中间室（活力）。我们将 Trk 融合基因 (TFG) 确定为该界面的关键组成部分，并表明其抑制作用使 ER 和 ERGIC 膜解偶联，导致 COPII 包被载体的各向同性扩散，降低货物分泌效率。我的第二个研究重点是了解其机制引导 MVE 的形成，MVE 使腔内囊泡 (ILV) 萌芽到其内部以隔离来自细胞质的膜相关货物。 MVE 与溶酶体的最终融合导致 ILV 及其相关蛋白的降解，通过控制 ILV 及其相关蛋白在肿瘤抑制中发挥关键作用信号受体的捕获和隔离。使用秀丽隐杆线虫，我们开发了一种新的、高度简化的、和遗传易处理系统来研究内体分选复合物的成分如何运输（ESCRT）机械使内吞货物能够在以下情况下移动到 MVE 和 ILV：一个完整的、正在发育的动物。我们未来的工作将继续利用不断发展的方法来进一步确定如何正确调节 COPII 介导的运输和 ESCRT 介导的 MVE 生物发生。更好的了解这些过程将有助于深入了解维持正常蛋白质的稳态控制细胞增殖和分化过程中分泌和内吞途径的运输。