Structural studies of respiratory syncytial virus

呼吸道合胞病毒的结构研究

• 批准号: 8512145
• 负责人: ELIZABETH R WRIGHT
• 金额: $ 23.4万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-15 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8512145
• 关键词: Address; Animals; Architecture; Area; Biochemical; Biological Models; Bronchiolitis; C-terminal; Cell membrane; Cells; Cellular Membrane; Cessation of life; Charge; Child; Cholesterol; Complex; Cryoelectron Microscopy; Crystallography; Detergents; Development; Elderly; Electron Microscope; Family; Freezing; Future; Generations; Glycoproteins; Goals; HIV-1; Hendra Virus; Human; Human Metapneumovirus; Human respiratory syncytial virus; Imaging technology; In Vitro; Individual; Infant; Infection; Influenza; Knowledge; Lipid Bilayers; Lipid Biochemistry; Location; Lung; Macromolecular Complexes; Measles; Measles virus; Mediating; Membrane; Membrane Glycoproteins; Membrane Microdomains; Methods; Microscopy; Molecular; Molecular Virology; Morphology; Myxovirus; N-terminal; Nipah Virus; Paramyxoviridae; Paramyxovirus; Pattern; Phase; Process; Production; Prophylactic treatment; Protein Binding; Protein Region; Proteins; Public Health; RNA Viruses; Recruitment Activity; Regulation; Resistance; Resolution; Respiratory syncytial virus; Ribonucleoproteins; Role; Site; Staging; Structural Protein; Structure; Technology; Tomogram; Translating; Vaccines; Viral; Viral Matrix Proteins; Viral Pneumonia; Viral Proteins; Viral Structural Proteins; Virion; Virus; Virus Assembly; Virus-like particle; base; driving force; electron tomography; glycoprotein G; glycoprotein structure; improved; in vivo; macromolecule; member; multiple myeloma M Protein; parainfluenza virus; particle; pathogen; public health relevance; reconstruction; research study; success; therapeutic vaccine; three dimensional structure; viral RNA; virus morphology

DESCRIPTION (provided by applicant): The matrix (M) protein is the major viral structural protein that regulates assembly and structure determination in members of the Para- and Ortho-myxoviridae families. The two families include numerous human pathogens such as human respiratory syncytial virus (hRSV), measles virus (MeV), human parainfluenza virus (hPIV), and the Influenza A, B, and C genera. The matrix protein initiates virus assembly by forming oligomeric sheets in specific regions along the host cell membrane. The M protein also orchestrates the incorporation of the viral glycoproteins and the ribonucleoprotein (RNP) into the evolving viral particles. Here we aim to determine two things: 1) the native structure and organization of RSV, which is an archetypical member of the Paramyxoviridae family; and 2) how the RSV M protein regulates virus assembly and determines virus morphology. To answer these questions, we are using the A2 strain of RSV that causes bronchiolitis, viral pneumonia, and death in young children, the elderly, and immuno-compromised individuals. We are analyzing RSV structure to build upon our previous structural studies of HIV-1 assembly and maturation, our ongoing structural analysis of native measles virus glycoprotein arrangements and interactions, and to develop RSV as a model system for future structural studies of virus assembly, virus organization, and the development of correlative microscopy methods. Together these studies will provide new information regarding the pleiomorphic structure of native RSV virions and the structural and functional role of the matrix protein in myxovirus assembly and morphology regulation. The two areas to be investigated in this project are: 1. Determine the native structure of RSV strain A2. Experiments will determine the natural structure of RSV through cryo-ET of frozen hydrated virions. Cryo-immuno-EM approaches and RSV RNA-specific probes will be used to define the placement and organizational patterns of the RSV structural proteins, F, G, SH, M, and the RNP complex during cryo-ET analysis of cryo-preserved virions. 2. Determine the structure of oligomeric RSV M in vivo and under in vitro assembly conditions. Experiments in this aim will determine the in vivo structure of RSV M in native virions and virus like particles (VLPs) through cryo-ET and sub-tomogram averaging. Cryo-ET, cryo-EM and helical reconstruction approaches will define the high-resolution structure of RSV M under in vitro assembly conditions.

描述（由申请人提供）：基质(M)蛋白是主要的病毒结构蛋白，调节准黏液病毒科和正黏液病毒科成员的组装和结构决定。这两个科包括许多人类病原体，如人类呼吸道合胞病毒（hRSV）、麻疹病毒（MeV）、人类副流感病毒（hPIV）以及甲型、乙型和丙型流感。基质蛋白通过在宿主细胞膜的特定区域形成寡聚物薄片来启动病毒组装。M蛋白还协调将病毒糖蛋白和核糖核蛋白（RNP）整合到进化的病毒颗粒中。本文旨在确定两件事：1)RSV的天然结构和组织，它是副粘病毒科的一个典型成员；2) RSV M蛋白如何调控病毒组装并决定病毒形态。为了回答这些问题，我们使用了引起毛细支气管炎、病毒性肺炎和幼儿、老年人和免疫功能受损个体死亡的RSV A2株。我们正在分析RSV的结构，以建立我们之前对HIV-1组装和成熟的结构研究，我们正在进行的对天然麻疹病毒糖蛋白排列和相互作用的结构分析，并将RSV作为未来病毒组装、病毒组织结构研究的模型系统，并开发相关的显微镜方法。这些研究将为RSV病毒载体的多形性结构以及基质蛋白在黏液病毒组装和形态调控中的结构和功能作用提供新的信息。本项目主要研究的两个方面是：1。确定RSV A2株的天然结构。实验将通过冷冻水合病毒粒子的冷冻- et来确定RSV的自然结构。冷冻免疫电镜方法和RSV rna特异性探针将用于确定RSV结构蛋白、F、G、SH、M和RNP复合物在冷冻et分析过程中的位置和组织模式。2. 在体内和体外组装条件下测定低聚RSV M的结构。本实验将通过冷冻et和亚断层扫描平均技术确定RSV M在天然病毒粒子和病毒样颗粒（VLPs）中的体内结构。Cryo-ET， cryo-EM和螺旋重建方法将在体外组装条件下定义RSV M的高分辨率结构。