MECHANISMS OF AP-3 FUNCTION IN VESICLE FORMATION AND GOLGI MATURATION

AP-3 在囊泡形成和高尔基体成熟中的功能机制

基本信息

批准号：
9815765
负责人：
Alexey Jarrell Merz
金额：
$ 48.95万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-12 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9815765
关键词：
Animal Model Architecture Binding Biochemical Biochemical Genetics Biochemistry Biological Biological Models Biological Process Bypass Capsid Cell physiology Cells Coated vesicle Communicable Diseases Complex Data Defect Destinations Disease Early Endosome Electron Microscopy Endocytosis Endosomes Epilepsy Eukaryotic Cell Event Fluorescence Microscopy Foundations Functional disorder Genetic Genetic Diseases Goals Golgi Apparatus HIV HIV-1 Hermanski-Pudlak Syndrome Human Impairment Lesion Link Lipids Location Lysosomes Membrane Microscopic Mission Modeling Molecular Morphology Multivesicular Body National Institute of General Medical Sciences Organelles Outcome Pathway interactions Pharmacology Play Prevention Process Proteins Proteomics Public Health Publishing Research Role Saccharomyces cerevisiae Saccharomycetales Site Sorting - Cell Movement Structure Surveys Syndrome System TFAP2A gene Testing Transcription Factor AP-1 Transport Vesicles Ubiquitination Vacuole Vesicle Vesicle Transport Pathway Viral Virus Assembly Work Yeasts base cofactor disease diagnosis follow-up functional genomics genetic approach imaging approach innovation late endosome novel prevent programs spatiotemporal trafficking trans-Golgi Network ubiquitin ligase ubiquitin-protein ligase yeast genetics

项目摘要

PROJECT SUMMARY The secretory pathway folds, assembles, and sorts membrane and secreted proteins and targets them to a variety of locations inside and outside the cell. The `Grand Central Station' of the cell is the trans Golgi network (TGN), an incredibly dynamic and complex structure that packages cargo into separate containers and dispatches them to their terminal destinations. The AP-3 trafficking complex sorts cargo from endosomes or the Golgi into containers bound for endolysosomal compartments, bypassing the endosomal multivesicular body and ESCRT pathway. AP-3 is needed for productive assembly of HIV and other viral capsids. Genetic lesions impairing AP-3 function cause multiple diseases, including Hermansky-Pudlak Syndrome type 2 and an epilepsy-like syndrome. Despite its importance, remarkably little is known about the mechanisms of AP-3 function. In this collaborative Project, we exploit published and unpublished genetic, cell biological, and biochemical advances, and employ state of the art imaging approaches, to dissect the mechanisms of AP-3 budding and the consequences of AP-3 function for the overall dynamics of the TGN, in the model organism Saccharomyces cerevisiae. In Aim 1, we build on proteomic surveys, functional genomic surveys, and follow-up analyses, wich identify new candidate AP-3 cofactors including a ubiquitin ligase. We will use these advances to test hypotheses about the assembly, maturation, and dispatch of AP-3 vesicles from the TGN. In Aim 2, we build on preliminary findings showing that disruption of AP-3 activity causes striking and unexpected global perturbations of Golgi architecture. We hypothesize, based on published and unpublished data, that distinct carriers exit the TGN in a programmed, sequential manner, with AP-3 carriers departing early. We will further test this model and dissect the molecular mechanisms though which the sequence of events is programmed. Together, these studies should establish new paradigms for AP-3 vesicle formation and for organelle maturation dynamics in the Golgi-endolysosomal system.

项目摘要分泌途径折叠，组装和分类膜和分泌的蛋白质，并将其靶向牢房内外的各种位置。电池的“大中央车站”是跨性别的高尔基网络（TGN）是一种令人难以置信的动态且复杂的结构，将货物包装到单独的容器中，并将其派往其终端目的地。 AP-3贩运复合物分类来自内体或高尔基体进入绑定到内溶性隔室的容器，绕过内体多囊体和escrt途径。艾滋病毒和其他病毒式衣壳的生产力组装需要AP-3。遗传病变损害AP-3功能引起多种疾病，包括2型Hermansky-Pudlak综合征和癫痫样综合征。尽管它的重要性很重要，但对AP-3功能的机制知之甚少。在这个合作项目中，我们利用出版和未发表的遗传，细胞生物学和生化进步，采用艺术成像方法的状态，以剖析AP-3萌芽的机制和 AP-3功能对TGN的总体动力学的后果，在模型生物体糖果中酿酒酵母。在AIM 1中，我们以蛋白质组学调查，功能性基因组调查和后续分析为基础识别包括泛素连接酶在内的新候选AP-3辅助因子。我们将使用这些进步来检验有关从TGN组装，成熟和调度AP-3囊泡的假设。在AIM 2中，我们以初步发现表明，AP-3活动的破坏会引起高尔基体体系结构的惊人和意外的全球扰动。我们根据已发表和未发表的数据假设，这是不同的载体以编程的，顺序的方式退出TGN，AP-3载体提早离开。我们将进一步测试虽然对事件的序列进行编程，并解剖分子机制。一起，这些研究应建立用于AP-3囊泡形成的新范例和Golgi-endolysosomal系统中的细胞器成熟动力学。