Structural and Functional Characterization of the Ebola Virus Replication Complex

埃博拉病毒复制复合物的结构和功能表征

• 批准号: 9149555
• 负责人: Gaya K. Amarasinghe
• 金额: $ 287.63万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-07 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9149555
• 关键词: Address; Adverse effects; Affect; Africa; Attenuated; Biochemistry; Biological Assay; Biological Testing; Biology; Case Study; Categories; Cessation of life; Collaborations; Complex; Cryoelectron Microscopy; DNA-Directed RNA Polymerase; Data Analyses; Development; Disease; Disease Outbreaks; Ebola virus; Elements; Ensure; Epidemic; Europe; Evaluation; Event; Filovirus; Genome; Goals; Health; Human; Human Resources; Immunology; In Vitro; Inflammation; Integration Host Factors; Knowledge; Light; Link; Manuscripts; Maps; Molecular; Monoclonal Antibodies; Nucleoproteins; Pathogenesis; Production; Program Research Project Grants; Proteins; Proteome; Proteomics; RNA; RNA Editing; RNA Interference; RNA Viruses; RNA chemical synthesis; RNA-Directed RNA Polymerase; Reagent; Recording of previous events; Regulation; Reporting; Research; Research Infrastructure; Research Personnel; Research Project Grants; Resolution; Resources; Signal Transduction; Specificity; Structure-Activity Relationship; Technical Expertise; Technology; Testing; Therapeutic; Training; Trans-Activators; Variant; Viral; Viral Hemorrhagic Fevers; Viral Proteins; Virus; Virus Replication; Work; X-Ray Crystallography; abstracting; antiviral immunity; base; biodefense; biophysical analysis; biosafety level 4 facility; feeding; genome-wide; global health; health economics; high throughput screening; in vivo; in vivo Model; innovation; insight; multidisciplinary; mutant; new therapeutic target; novel; pathogen; programs; structural biology; synthetic antibodies; therapeutic development; viral RNA; virology

Abstract: Project Overview for Structural and Functional Studies of Ebola Virus RNA synthesis Ebolaviruses (EBOVs) are non-segmented negative-strand RNA viruses (NNSVs) and Category A Priority biodefense pathogens that cause frequent lethal hemorrhagic fever, including the devastating and ongoing outbreak in West Africa. Approved anti-EBOV therapeutics are lacking. EBOV replicates with high efficiency in vivo while effectively evading host innate antiviral immunity. The unrestrained replication and associated inflammation leads to death. A complete understanding of EBOV pathogenesis therefore requires understanding how the viral RNA dependent RNA polymerase (RDRP) complex interacts with host factors. The overarching goal of this program project proposal is to understand the mechanisms of RDRP function by defining the interactions among its components and with the host proteome and by defining signals that regulate its function. Toward this goal Project 1 will identify cis- and trans-acting factors that impact EBOV RDRP activity. Project 2 will define a structural basis for the EBOV RDRP complex, identify regulatory mechanisms, and determine species and strain specificities, and Project 3 will use genome-wide RNAi and proteomic strategies to identify host factors that modulate RDRP function. Project 1 provides functional context into which a subset of Project 3 “hits” can be interpreted, while Project 3 will likely identify host factors relevant to the findings of Project 1. Project 2 will provide a structural framework that will be used to integrate findings from both Projects 1 and 3. Together, these studies will provide mechanistic insights into how protein-protein and protein-vRNA interactions control EBOV RDRP function. Research projects are supported by an Administrative Core, a Protein Production and Protein Interaction Core, and critically, a Biosafety Level 4 (BSL4) Core. Core B will generate unique reagents, including monoclonal and synthetic antibodies. Core C (BSL4) will provide unique capabilities to obtain materials from and perform tests using wild type and mutant EBOVs as well as pathogenesis evaluation. The work will be performed by highly productive and collaborative investigators with expertise in every aspect of the proposed studies, including biochemistry, EBOV pathogenesis, high throughput screening and data analysis, immunology, proteomics, structural biology, and virology. These studies will provide unprecedented insights into the components of the EBOV RDRP and in its interaction with the host as well as species and strain differences that affect the RDRP complex. We also expect to identify specific viral and host factor interactions as novel therapeutic targets.

摘要：用于埃博拉病毒RNA合成结构和功能研究的项目概述 埃博氏病毒（EBOV）是非细分的负链RNA病毒（NNSV）和类别的优先级 经常引起致命的出血热的生物延伸病原体，包括毁灭性和正在进行的 西非爆发。缺乏批准的抗EBOV疗法。 EBOV以高效率复制 体内有效地逃避了宿主先天的抗病毒免疫。不受约束的复制和相关的 炎症导致死亡。因此，完全了解EBOV发病机理需要 了解病毒RNA依赖性RNA聚合酶（RDRP）复合物如何与宿主因子相互作用。 该计划项目建议的总体目标是通过 定义其组件之间以及与宿主蛋白质组之间的相互作用，并通过定义信号 调节其功能。朝着这个目标项目1识别影响EBOV的顺式和跨性别因素 RDRP活动。项目2将定义EBOV RDRP复合物的结构基础，确定调节 机制，并确定规格和应变规格，项目3将使用全基因组RNAi和 蛋白质组学策略以识别调节RDRP功能的宿主因素。项目1提供功能上下文 项目3的子集可以解释“命中”，而项目3可能会确定宿主因素相关 项目1。项目2的发现将提供一个结构框架，该框架将用于整合发现 从两个项目1和3中。一起，这些研究将提供有关蛋白质蛋白质的机械见解 和蛋白质-VRNA相互作用控制EBOV RDRP功能。研究项目得到 行政核心，蛋白质的产生和蛋白质相互作用核心，以及批判性的生物安全水平4 （BSL4）核心。核B将产生独特的试剂，包括单克隆和合成抗体。核心c （BSL4）将提供独特的功能，以获取使用野生型和突变体的材料并进行测试 EBOV以及发病机理评估。这项工作将由高产和协作进行 在拟议研究的各个方面都有专业知识的研究人员，包括生物化学，EBOV 发病机理，高通量筛查和数据分析，免疫学，蛋白质组学，结构生物学和 病毒学。这些研究将为EBOV RDRP的组成部分提供前所未有的见解 与宿主以及物种的相互作用以及影响RDRP复合物的应变差异。我们也是 期望将特定的病毒和宿主因子相互作用确定为新的治疗靶标。