Biochemical and structural mechanisms at the filoviral-host interface

丝状病毒-宿主界面的生化和结构机制

• 批准号: 10555055
• 负责人: Gaya K. Amarasinghe
• 金额: $ 111.25万
• 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 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10555055
• 关键词: Actins; Address; Africa; Antiviral Agents; Biochemical; Biochemistry; Biology; COVID-19 pandemic; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Communicable Diseases; Complement; Complex; Core Protein; Cryo-electron tomography; Cryoelectron Microscopy; Cytoskeleton; Data; Development; Disease; Disease Outbreaks; Ebola virus; Event; Family; Filovirus; Genetic Transcription; Genome; Health; Human; Hybrids; Immune; Immune response; Infection; Integration Host Factors; Knowledge; Length; Low-Density Lipoproteins; Marburgvirus; Mass Spectrum Analysis; Methods; Molecular; Nature; Nucleocapsid; Nucleocapsid Proteins; Nucleoproteins; Open Reading Frames; Pathway interactions; Physical condensation; Play; Polymerase; Positioning Attribute; Post-Translational Protein Processing; Process; Productivity; Protein-Protein Interaction Map; Proteins; Proteomics; Publications; Publishing; RNA; RNA chemical synthesis; RNA-Directed RNA Polymerase; Regulation; Research Personnel; Research Project Grants; Resolution; Resources; Roentgen Rays; Role; Signal Transduction; Small Interfering RNA; Structure; Therapeutic; Time; Viral; Viral Proteins; Virus; Virus Diseases; Virus Replication; Work; X-Ray Crystallography; enzyme activity; global health; innovation; insight; multidisciplinary; multiple omics; novel; nuclease; particle; protein protein interaction; receptor; structural biology; therapeutic target; tool; vaccine candidate; viral RNA

RP01 Project Summary/Abstract for Biochemical and structural mechanisms at the filoviral-host interface The recent filoviral outbreaks, including the 2013-2016 Ebola virus (EBOV) outbreak in West Africa that introduced the virus to other continents, highlight the imminent threat to global health posed by filoviruses and the urgent need for basic and translational efforts. The current COVID-19 pandemic further illustrates the significance of understanding infectious diseases. While recent efforts to develop countermeasures have resulted in vaccine candidates and some therapeutics, filoviruses remain a considerable threat to human health and key questions are still outstanding. There are substantial gaps in our understanding of host-viral interactions that contribute to disease, including how viral proteins limit host responses and assemble into infectious particles. Work in Research Project 1 (RP01) will use proteomic, biochemical, and structural methods to identify and validate molecular mechanisms at the host-viral interface in cellular networks to address this gap. We will address these longstanding mechanistic questions using biochemical and hybrid structural methods, including mass spectrometry, NMR, X-ray crystallography, small angle X-ray scattering (SAXS), and cryoelectron microscopy (cryo-EM), cryoelectron tomography (cryo-TM) to characterize filoviral nucleocapsid (NC) interactions and to define high impact filoviral-host factor interactions modulating PTMs that impact the viral replication cycle, viral NC assembly, and egress. Our strong publication record, preliminary results from ongoing studies, and access to unique facilities and resources support these efforts. We are uniquely positioned due to our highly productive and collaborative team with complementary expertise and a prior record of co-authored studies with investigators in RP02, RP03, Core B, and Core C. Our Aims are: Aim 1. Determine the structural basis and dynamics of the filoviral nucleocapsid (NC) and define NC-host interactions; Aim 2. Develop Protein-protein interaction (PPI) maps that include post-translational modifications (PTMs) from virally infected cells for EBOV and MARV; and Aim 3. Define the molecular mechanisms for PPIs that contribute to filoviral infection defined by this project, RP02 and RP03. At the completion, we expect to define filoviral interaction with host factors that contribute to filoviral infection. Our arsenal of tools from mass spectrometry, biochemistry, and structural biology, together with work from the Research Projects and Scientific Cores within the PPG, enable us to clearly define each contribution to filoviral infection and identify new targets for antivirals.

RP 01丝状病毒宿主的生化和结构机制的项目总结/摘要 接口 最近爆发的丝状病毒疫情，包括2013-2016年西非爆发的埃博拉病毒（EBOV）， 将该病毒引入其他大陆，强调丝状病毒对全球健康构成的迫在眉睫的威胁， 迫切需要基本的和转化的努力。当前的COVID-19疫情进一步说明了 了解传染病的重要性。虽然最近为制定对策所作的努力 尽管丝状病毒已经产生了疫苗候选物和一些治疗剂，但丝状病毒仍然对人类构成相当大的威胁。 卫生和关键问题仍然悬而未决。在我们对宿主病毒的理解上， 导致疾病的相互作用，包括病毒蛋白如何限制宿主反应并组装成 传染性粒子研究项目1（RP 01）的工作将使用蛋白质组学，生物化学和结构 鉴定和验证细胞网络中宿主-病毒界面的分子机制的方法， 弥补这一差距。我们将利用生物化学和混合动力学来解决这些长期存在的机械问题。 结构方法，包括质谱、NMR、X射线晶体学、小角X射线散射 （SAXS）和冷冻电子显微镜（cryo-EM）、冷冻电子断层扫描（cryo-TM）来表征丝状病毒 核衣壳（NC）相互作用，并定义调节PTM的高影响丝状病毒-宿主因子相互作用， 影响病毒复制周期、病毒NC组装和外出。我们强大的出版记录，初步 正在进行的研究的结果以及获得独特设施和资源的机会支持这些努力。我们 独特的定位，由于我们的高效和协作的团队与互补的专业知识和 RP 02、RP 03、核心B和核心C中与研究者共同撰写研究的既往记录。我们的目标：Aim 1.确定丝状病毒核衣壳（NC）的结构基础和动力学，并定义NC宿主 目标2.开发包括翻译后修饰的蛋白质相互作用（PPI）图谱 用于EBOV和MARV的来自病毒感染细胞的PTM;和Aim 3。定义PPI的分子机制 导致本项目定义的丝状病毒感染，RP 02和RP 03。在完成时，我们希望 定义丝状病毒与导致丝状病毒感染的宿主因子的相互作用。我们的武器库从大众 光谱，生物化学和结构生物学，以及研究项目的工作， PPG中的科学核心使我们能够清楚地定义对丝状病毒感染的每一种贡献，并确定 抗病毒药物的新靶点