Biochemical and structural mechanisms at the filoviral-host interface

RP01项目摘要/丝状病毒宿主的生化和结构机制摘要 接口 最近的丝状病毒疫情，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，强调丝状病毒和丝状病毒对全球健康构成的迫在眉睫的威胁 迫切需要基本的和翻译的努力。目前的新冠肺炎大流行进一步说明 认识传染病的重要意义。虽然最近制定对策的努力已经 由于出现了候选疫苗和一些治疗方法，丝状病毒仍然对人类构成相当大的威胁 健康和关键问题仍然悬而未决。我们对宿主病毒的理解有很大的差距 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 有感染性的颗粒。正在研究的项目1(RP01)将使用蛋白质组、生化和结构 在细胞网络中识别和验证宿主-病毒界面的分子机制的方法 弥补这一差距。我们将使用生化和杂交技术来解决这些长期存在的机械问题 结构方法，包括质谱学、核磁共振、X射线结晶学、小角X射线散射 (SAXS)，以及冷冻电子显微镜(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的高影响力丝状病毒-宿主因子相互作用 影响病毒复制周期、病毒NC组装和出口。我们良好的出版记录，初步 正在进行的研究的结果，以及获得独特的设施和资源，都支持这些努力。我们是 独一无二的定位，得益于我们高效、协作的团队，他们拥有互补的专业知识和 在RP02、RP03、核心B和核心C中与研究人员共同撰写研究的先前记录。我们的目标是：AIM 1.确定丝状病毒核衣壳(NC)的结构基础和动力学，并确定NC宿主 相互作用；目标2.开发包括翻译后修饰的蛋白质-蛋白质相互作用(PPI)图 从EBOV和MARV的病毒感染细胞中提取(PTM)；以及目的3.定义PPI的分子机制 该项目RP02和RP03定义的丝状病毒感染。在完成后，我们预计将 定义丝状病毒与宿主因素的相互作用，这些因素有助于丝状病毒感染。我们来自麻省理工学院的工具库 光谱学、生物化学和结构生物学，以及研究项目和 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。

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.