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（由申请方提供）：基质（M）蛋白是调节帕拉科和邻黏病毒科成员组装和结构确定的主要病毒结构蛋白。这两个科包括许多人类病原体，如人呼吸道合胞病毒（hRSV）、麻疹病毒（MeV）、人副流感病毒（hPIV）和甲型、B和丙型流感病毒属。基质蛋白通过在宿主细胞膜的特定区域沿着形成寡聚片层来启动病毒装配。M蛋白还协调病毒糖蛋白和核糖核蛋白（RNP）掺入进化的病毒颗粒中。在这里，我们的目标是确定两件事：1）RSV的天然结构和组织，这是副粘病毒科的原型成员;和2）RSV M蛋白如何调节病毒装配和决定病毒形态。为了回答这些问题，我们正在使用RSV的A2毒株，该毒株可引起毛细支气管炎、病毒性肺炎和幼儿、老年人和免疫功能低下者的死亡。我们正在分析RSV结构，建立在我们以前的HIV-1组装和成熟的结构研究，我们正在进行的结构分析天然麻疹病毒糖蛋白的安排和相互作用，并开发RSV作为未来的病毒组装，病毒组织的结构研究的模型系统，和相关的显微镜方法的发展。总之，这些研究将提供新的信息，天然RSV病毒粒子的多形结构和结构和功能的作用，在粘病毒组装和形态调控的基质蛋白。本项目将研究的两个领域是：1。测定RSV毒株A2的天然结构。实验将通过冷冻水合病毒粒子的冷冻ET确定RSV的天然结构。冷冻免疫EM方法和RSV RNA特异性探针将用于在冷冻保存病毒体的冷冻ET分析期间确定RSV结构蛋白F、G、SH、M和RNP复合物的位置和组织模式。2.在体内和体外组装条件下测定寡聚RSV M的结构。为此目的的实验将通过冷冻ET和亚断层图像平均来确定天然病毒体和病毒样颗粒（VLP）中RSV M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确定RSV M的高分辨率结构。