Molecular biology of viral capsids

病毒衣壳的分子生物学

基本信息

批准号：
7976300
负责人：
Cheng C Kao
金额：
$ 34.18万
依托单位：
TRUSTEES OF INDIANA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-01 至 2014-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7976300
关键词：
Affinity Chromatography Architecture Arginine Basic Amino Acids Biochemical Biological Assay Biological Models Brome mosaic virus Buffers Capsid Capsid Proteins Cells Charge Complex Confocal Microscopy Cryoelectron Microscopy Detection Electron Microscopy Encapsulated Genome Goals Immune Immune response Infection Label Lead Length Location Maps Mediating Membrane Microtomy Modeling Modification Molecular Biology Molecular Genetics N-terminal Penetration Peptides Plant Model Plant Viruses Plants Primer Extension Process Production Proteins RNA RNA Conformation RNA Virus Infections RNA Viruses RNA chemical synthesis RNA replication RNA-Binding Proteins RNA-Protein Interaction Regulation Reporter Role Shapes Signal Transduction Solutions Structure Tail Tissues Trans-Activators Transducers Translation Process Translations Vacuole Viral Viral Genome Virion Virus Virus Diseases Work crosslink design genetic analysis improved insight mutant nanoGold particle public health relevance receptor reconstruction replicase research study viral RNA

项目摘要

DESCRIPTION (provided by applicant): Viral capsid proteins are important regulators of viral processes from translation, to RNA replication, to RNA packaging, and the modulation of cellular innate immune responses. In viruses with segmented genomes that are packaged into separate virions, the capsid protein must coordinate the activities for each of the RNAs. The overall goal of this project is to better understand the early steps in an RNA virus infection. That is, how does the capsid interact with the viral RNAs within the virion and facilitate entry into cells? Brome mosaic virus (BMV) will be the model virus for this study. BMV is a nonenveloped segmented positive-strand RNA virus that has served as a model system with notable contributions to the cis- and trans-acting factors, the formation of the replicase complex, and the mechanism of viral RNA-dependent RNA synthesis. Preliminary results showed that an arginine-rich motif of the BMV capsid contributes to the selective encapsidation of BMV RNAs and acts as a cell-penetration signal. The latter set of observations led to the hypothesis that plant viruses gain entry into cells through a protein transduction mechanism. This project contains two aims, the first will seek to understand the architecture of the virion through a combination of cryoelectron microscopy and single particle reconstruction, biochemical mapping of RNA conformation and RNA-capsid interaction, and a molecular genetic analysis of how the interactions will influence the infection process. The second aim seeks to elucidate the mechanism of cell entry of the cell-penetration peptides and the viral capsid using fluorescently-labeled peptides and viruses, confocal microscopy and electron microscopy. The work will lead to improved understanding of the viral infection process. PUBLIC HEALTH RELEVANCE: Viral capsid proteins have many essential roles during a virus infection, from the regulation of translation to the protection of the viral genome against cellular innate immune detection. This proposal contains a comprehensive set of experiments to examine how the capsid protein of a plant-infecting RNA virus interacts with the viral RNA, reconfigures the shape of the RNA during encapsidation, and mediates entry into cells. The information gained from the proposed studies will contribute new insights to virus architecture, protein-RNA interactions, and how viruses gain entry into the cell.

描述（由申请人提供）：病毒式衣壳蛋白是病毒过程的重要调节剂，从翻译，RNA复制，再到RNA包装以及细胞先天免疫反应的调节。在包装成单独的病毒体的分段基因组的病毒中，衣壳蛋白必须协调每个RNA的活性。该项目的总体目标是更好地了解RNA病毒感染的早期步骤。也就是说，衣壳如何与病毒体内的病毒RNA相互作用并促进进入细胞？ Brome Mosaic病毒（BMV）将是这项研究的模型病毒。 BMV是一种非发达的分段阳性RNA病毒，已充当模型系统，对顺式和反式作用因子，复制酶复合物的形成以及病毒RNA依赖性RNA合成的机制有显着贡献。初步结果表明，BMV衣壳的精氨酸基序有助于BMV RNA的选择性封装，并充当细胞渗透信号。后者的观察结果导致了以下假设：植物病毒通过蛋白质转导机制进入细胞。该项目包含两个目的，第一个将寻求通过冷冻电子显微镜和单个颗粒重建，RNA构象和RNA粘合膜相互作用的生化映射以及对相互作用如何影响感染过程的分子遗传分析的结合来理解病毒粒子的结构。第二个目的旨在阐明使用荧光标记的肽和病毒，共聚焦显微镜和电子显微镜使用荧光标记的肽和病毒的细胞渗透肽和病毒衣壳的机理。这项工作将导致人们对病毒感染过程的了解。公共卫生相关性：病毒式衣壳蛋白在病毒感染中具有许多基本作用，从调节到保护病毒基因组免受细胞先天免疫检测的保护。该建议包含一组全面的实验，以检查植物感染RNA病毒的衣壳蛋白如何与病毒RNA相互作用，重新配置封装过程中RNA的形状，并介导进入细胞。从拟议的研究中获得的信息将为病毒架构，蛋白-RNA相互作用以及病毒如何进入细胞提供新的见解。