Interplay of the HIV-1 Env cytoplasmic tail, Gag-MA, and membrane: resolving molecular detail and blocking assembly

HIV-1 Env 胞质尾部、Gag-MA 和膜的相互作用：解析分子细节并阻断组装

• 批准号: 10772333
• 负责人: Cynthia Ann Derdeyn
• 金额: $ 82.76万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-03 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10772333
• 关键词: Amino Acid Substitution; Antiviral Agents; Binding; Biochemical; Biochemistry; Biological Assay; Cell Membrane Permeability; Cells; Charge; Coupled; Cryo-electron tomography; Cyclic Peptides; Cytoplasmic Tail; Cytoplasmic matrix; Data; Drug Design; Epitopes; Evaluation; Generations; Goals; Grain; HIV; HIV envelope protein; HIV-1; Image; In Vitro; Length; Life; Lipids; Lytic; Mechanics; Membrane; Membrane Lipids; Modeling; Molecular; Molecular Cloning; Mutation; Nature; Peptides; Phosphatidylinositol 4,5-Diphosphate; Positioning Attribute; Production; Protein Engineering; Recombinants; Reporting; Resolution; Role; Structure; Testing; Time; Transmembrane Domain; Variant; Viral; Viral Physiology; Virion; Virus; Work; cell type; cost; density; design; env Gene Products; fitness; flexibility; in silico; in situ imaging; inhibitor; innovation; insight; molecular dynamics; multi-scale modeling; multidisciplinary; novel; novel therapeutics; particle; simulation; synergism; transmission process; virology

PROJECT SUMMARY/ABSTRACT The goal of this innovative project is to bring together multidisciplinary expertise to dissect the functional, structural, and dynamic nature of interactions between the HIV-1 envelope (Env) cytoplasmic tail (CT), the underlying Gag matrix (MA) lattice, and the lipid containing membrane in the context of native virions. We will utilize state-of-the-art cryo-electron tomography (cryo-ET), molecular dynamics (MD) simulations, modeling, and in vitro studies to produce a comprehensive rendition of how interactions between these key determinants of assembly and infectivity are orchestrated. Our studies are based on recent cryo-ET data in which the positioning of Env CT and MA interactions differed from previous models. In Aim 1, we will harness the power of cryo-ET to advance the resolution of Env CT-MA interactions to facilitate further interrogation of the mechanics of Env CT- MA-membrane interactions. In Aim 2, we will generate atomistic models of Env CT-MA in the context of the viral membrane using high resolution structures and cryo-ET density, and strategic mutations and truncations of Env CT and MA. In Aim 3, we will exploit conserved structural features to design novel antiviral agents comprised of membrane-permeable cyclic peptide inhibitors. These Aims are supported by comprehensive biochemical and virologic approaches and will use a well characterized clade C transmitted/founder (T/F) HIV-1 infectious molecular clone that is fully representative of global isolates and presents fully matched Env and Gag components, which distinguishes our work. Our studies will reveal in unprecedented detail how Env CT, MA, and membrane lipids engage one another, with all components in near native context. These powerful approaches will produce an integrated model, including flexible regions that are difficult to resolve, to bring the complete Env- MA-membrane machinery to life.

项目摘要/摘要 这个创新项目的目标是将多学科的专业知识结合在一起，剖析 HIV-1包膜(Env)细胞质尾部(CT)、HIV-1包膜(Env)和胞浆尾部(CT)之间相互作用的结构和动态性质 在天然病毒粒子的背景下，下面的GAG基质(MA)晶格和含脂的膜。我们会 利用最先进的冷冻电子断层扫描(CRYO-ET)、分子动力学(MD)模拟、建模和 体外研究，以产生一个全面的再现之间的这些关键决定因素之间的互动 集合性和感染性是精心策划的。我们的研究是基于最近的低温数据，其中定位 环境、CT和MA的交互作用不同于以前的模型。在目标1中，我们将利用冷冻的力量 提高环境CT-MA相互作用的分辨率，以促进对环境CT-MA力学的进一步询问 MA-膜相互作用。在目标2中，我们将在病毒环境中生成Env CT-MA的原子模型 利用高分辨率结构和低温密度的膜，以及环境的战略突变和截断 CT和MA。在目标3中，我们将利用保守的结构特征来设计新的抗病毒药物，包括 膜透性环肽抑制剂。这些目标得到了全面的生化和 病毒学接近并将使用具有良好特征的C分支传播/创始人(T/F)HIV-1感染性 完全代表全球分离株并呈现完全匹配的env和gag的分子克隆 组件，这使我们的工作与众不同。我们的研究将以前所未有的细节揭示Env CT、MA和 膜脂相互作用，所有成分都处于接近天然的环境中。这些强大的方法 将产生一个集成的模式，包括难以解决的灵活区域，以带来完整的环境- 马膜机械要活起来。