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Hsp70 as Dispatcher of Virus Assembly and Trafficking

Hsp70 作为病毒组装和贩运的调度者

基本信息

批准号：
7272045
负责人：
VALERIAN V. DOLJA
金额：
$ 23.14万
依托单位：
OREGON STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-09-01 至 2009-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7272045
关键词：
ATP phosphohydrolase Address Animals Architecture Atomic Force Microscopy Beet yellows virus Biochemical Biochemical Genetics Biological Biological Assay Capsid Carrier Proteins Cells Code Collection Cytoskeleton Devices Dissection Docking Electron Microscopy Electrons Environment Experimental Models Funding Genetic Homologous Gene Human In Situ In Vitro Infection Investigation Life Life Cycle Stages Mass Screening Mass Spectrum Analysis Microfilaments Microscopy Microtubules Modeling Modification Molecular Molecular Analysis Molecular Chaperones Movement Organelles Organism Pathway interactions Plant Viruses Plants Plasmodesmata Process Proteins RNA Viruses Reagent Recruitment Activity Relative (related person)Reporter Research Role Role playing therapy Site System Tail Testing Time Variant Vascular System Viral Viral Proteins Virion Virus Virus Assembly Virus Diseases Virus-Cell Membrane Interaction Yeasts cell assembly cell motility design in vivo innovation intercellular communication intracellular protein transport mutant protein function protein transport tool trafficking virus host interaction yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Infection cycles of the human, animal, and plant viruses include processes of virion assembly and dissemination within the infected organism. These processes require functions of the specialized viral capsid and transport proteins, as well as cellular proteins and organelles recruited by virus. The functional coordination among viral and host components during virion formation and spread are understood relatively poorly. The beet yellows virus (BYV) model was developed during this project to reveal interactions between the viral and cell proteins using innovative biochemical, genetic and cell biological approaches. The BYV homolog of Hsp70 molecular chaperones (Hsp70h) was found to integrate principal functions of virion assembly and transport with modifications of the cell architecture. The BYV model provides unique advantages for investigation of these processes, including ability to visualize virus-host interactions within the live infected cells. Two Specific Aims are in the focus of this proposal. Aim 1 is designed to determine the molecular architecture of the BYV virions. These virions possess the tails, which serve as specialized movement devices empowered by Hsp70h. The topology of the tail proteins will be resolved using electron and atomic force microscopy. The Hsp70h-guided mechanism of the tail assembly will be dissected using available in planta model. Aim 2 will address the interactions between Hsp70h and the host cell. The hypothesis stating that Hsp70h interacts with host proteins to remodel cellular environment and facilitate virus trafficking in association with the cytoskeletal motility systems will be tested. Using BYV variants that express organelle-targeted fluorescent reporters, real-time changes in cell architecture during virus infection will be examined. Host proteins that interact with Hsp70h will be identified using yeast two-hybrid screens and mass spectrometry analyses. Functional significance of the observed interactions will be assessed using genetic strategies targeting both Hsp70h and host proteins.

描述(由申请人提供)：人类、动物和植物病毒的感染周期包括病毒粒子在受感染有机体内组装和传播的过程。这些过程需要专门的病毒衣壳和运输蛋白以及病毒招募的细胞蛋白和细胞器的功能。病毒粒子形成和传播过程中病毒和宿主成分之间的功能协调相对较少。甜菜黄化病毒(BYV)模型是在该项目期间开发的，旨在利用创新的生化、遗传和细胞生物学方法揭示病毒和细胞蛋白之间的相互作用。Hsp70分子伴侣的BYV同源物(Hsp70h)将病毒粒子组装和运输的主要功能与细胞结构的改变结合在一起。BYV模型为研究这些过程提供了独特的优势，包括可视化活感染细胞内病毒与宿主的相互作用的能力。这项提案的重点是两个具体目标。目的1是为了确定BYV病毒粒子的分子结构。这些病毒粒子拥有尾巴，尾巴是由Hsp70h提供动力的专门移动设备。尾部蛋白质的拓扑结构将使用电子显微镜和原子力显微镜进行解析。尾部组件的Hsp70h导向机构将使用Planta模型中的现有模型进行解剖。目标2将解决Hsp70h与宿主细胞之间的相互作用。假设Hsp70h与宿主蛋白相互作用，以重塑细胞环境，并促进与细胞骨架运动系统相关的病毒运输，这一假设将得到测试。使用表达细胞器靶向荧光报告的BYV变体，将检测病毒感染期间细胞结构的实时变化。与Hsp70h相互作用的宿主蛋白将使用酵母双杂交筛选和质谱分析来鉴定。将使用针对Hsp70h和宿主蛋白的遗传策略来评估观察到的相互作用的功能意义。