Mechanisms of actin cytoskeleton modulation by Pneumoviruses

肺病毒调节肌动蛋白细胞骨架的机制

基本信息

批准号：
10160770
负责人：
Rebecca E. Dutch
金额：
$ 60.38万
依托单位：
UNIVERSITY OF KENTUCKY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-11 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10160770
关键词：
Actins Address Affect Affinity Chromatography Bacteria Binding Biochemical Biochemistry Biological Biological Assay Biological Models Biophysics Bronchiolitis Calorimetry Cell physiology Cells Cellular Morphology Cellular biology Childhood Co-Immunoprecipitations Complex Coupled Cytoskeletal Proteins Cytoskeleton Data Deuterium Disease Dyes Elderly Elements Family Fluorescent in Situ Hybridization Human Human Metapneumovirus Hybrids Hydrogen Immunocompromised Host In Vitro Individual Infection Life Cycle Stages Ligation Lower Respiratory Tract Infection Mass Spectrum Analysis Mediating Microscopy Modification Molecular Molecular Sieve Chromatography Mutation Analysis NMR Spectroscopy Nature Nucleocapsid Pathogenesis Phosphoproteins Physiological Play Pneumovirus Pneumovirus Infections Population Process Proteins RNA RNA Viruses Respiratory Syncytial Virus Infections Respiratory Tract Infections Respiratory syncytial virus Role Structure System Testing Time Titrations Viral Viral Proteins Virus Virus Diseases Virus Replication X-Ray Crystallography airway epithelium cell motility cellular targeting effective therapy human pathogen insight intercellular communication light scattering member mutant new therapeutic target novel pathogenic virus polymerization respiratory pathogen structural biology therapeutic development therapeutic target viral RNA virology

项目摘要

The actin cytoskeleton controls key cellular processes, including cell morphology, motility, and intercellular communication. Many viral pathogens exploit the actin cytoskeletal machinery by a variety of unique mechanisms to facilitate viral infection, replication, and egress. Currently, the molecular underpinnings of these viral mechanisms remain poorly defined for a majority of viruses, including members of the Pneumoviridae family. Human respiratory syncytial virus (RSV) and human metapneumovirus (HMPV) are non-segmented negative strand RNA viruses that are members of the recently created Pneumoviridae family. RSV and HMPV cause significant disease, including bronchiolitis and lower respiratory tract infection in the pediatric population, elderly, and immunocompromised individuals. Recent studies from our groups and others have implicated interactions between actin or actin-interacting proteins and specific pneumovirus proteins in replication and viral spread; however, the mechanisms and cellular targets required for facilitating actin rearrangements remain unknown. Our overall hypothesis is that a cascade of specific interactions between pneumovirus proteins and the actin cytoskeleton regulates key steps in viral infection, including formation of novel structures critical for virus cell-to-cell spread. Our collaborative team, with expertise in virology, cell biology, biochemistry, and structural biology, will test our hypothesis through three specific aims. First, we will dissect the interactions between RSV and HMPV matrix (M) proteins and phosphoproteins (P) that regulate the actin cytoskeleton within an infected cell. Second, we will determine the biochemical and structural basis for HMPV phosphoprotein/matrix/actin complex and RSV P/M/actin complex. Finally, we will determine the mechanisms of intercellular extension formation, stabilization, and utilization in viral spread. At the completion of these studies, we expect to define molecular mechanisms by which RSV and HMPV interact with and modulate the host actin cytoskeleton during virus replication, define key elements needed for this modulation, and understand the molecular details of cell-to-cell spread through intercellular extensions, thus providing critical new insights that may be applicable in multiple viral systems.

肌动蛋白细胞骨架控制关键细胞过程，包括细胞形态，运动和细胞间沟通。许多病毒病原体通过多种独特的促进病毒感染，复制和出口的机制。目前，这些分子基础大多数病毒的病毒机制仍然很差，包括肺炎病毒的成员家庭。人呼吸综合病毒（RSV）和人元病毒（HMPV）是非细分的负链RNA病毒是最近创建的肺炎病毒家族的成员。 RSV和HMPV 引起严重疾病，包括小儿人群中的细支气管炎和下呼吸道感染，老年人和免疫功能低下的个体。我们小组和其他人的最新研究牵涉到在复制和病毒传播；但是，促进肌动蛋白重排需要的机制和细胞靶仍然未知。我们的总体假设是肺炎病毒之间的一系列特定相互作用蛋白质和肌动蛋白细胞骨架调节病毒感染的关键步骤，包括形成新型对于病毒细胞到细胞扩散至关重要的结构。我们的合作团队，具有病毒学专业知识，细胞的专业知识生物学，生物化学和结构生物学将通过三个特定目标检验我们的假设。首先，我们会的剖析RSV和HMPV基质（M）蛋白和磷蛋白（P）之间的相互作用，以调节感染细胞内的肌动蛋白细胞骨架。其次，我们将确定生化和结构基础 HMPV磷蛋白/基质/肌动蛋白复合物和RSV P/M/肌动蛋白复合物。最后，我们将确定病毒扩散中细胞间延伸形成，稳定和利用的机制。完成在这些研究中，我们期望定义RSV和HMPV与之相互作用的分子机制在病毒复制过程中调节宿主肌动蛋白细胞骨架，定义此调制所需的关键元素，并了解细胞对细胞细胞间扩散的分子细节，从而提供可能适用于多个病毒系统的关键新见解。