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.

项目概要/摘要 这个创新项目的目标是汇集多学科专业知识来剖析功能、 HIV-1 包膜 (Env) 细胞质尾 (CT) 之间相互作用的结构和动态性质 底层的 Gag 矩阵 (MA) 晶格，以及天然病毒颗粒背景下的含脂质膜。我们将 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，全面解释这些关键决定因素之间的相互作用 组装和感染力是精心策划的。我们的研究基于最近的冷冻电子断层扫描数据，其中定位 Env CT 和 MA 相互作用的结果与以前的模型不同。在目标 1 中，我们将利用冷冻 ET 的力量 推进 Env CT-MA 相互作用的分辨率，以促进进一步探究 Env CT-MA 的力学原理 MA-膜相互作用。在目标 2 中，我们将在病毒背景下生成 Env CT-MA 的原子模型 使用高分辨率结构和冷冻电子断层密度以及 Env 的策略突变和截断的膜 CT 和 MA。在目标 3 中，我们将利用保守的结构特征来设计新型抗病毒药物，其中包括 膜渗透性环肽抑制剂。这些目标得到全面的生化和 病毒学方法，并将使用特征明确的 C 分支传播/创始人 (T/F) HIV-1 传染性 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 膜脂相互结合，所有成分都处于接近天然的环境中。这些强有力的方法 将产生一个集成模型，包括难以解决的灵活区域，以带来完整的Env- MA-膜机械的生命力。