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汇集多学科专业知识，以剖析功能， HIV-1包膜（Env）细胞质尾巴（CT）之间相互作用的结构和动态性质， 在天然病毒体的背景下，基础插入式矩阵（MA）晶格和含有膜的脂质。我们将 利用最先进的冷冻电子断层扫描（冷冻-ET），分子动力学（MD）模拟，建模和 体外研究，以对这些关键决定因素之间的相互作用的全面演绎 组装和感染力是精心策划的。我们的研究基于最新的冷冻数据数据 ENV CT和MA相互作用的不同与以前的模型不同。在AIM 1中，我们将利用Cryo-Et的力量 推进ENV CT-MA相互作用的分辨率，以促进对Env CT-机制的进一步审问 MA膜相互作用。在AIM 2中，我们将在病毒的背景下生成Env CT-MA的原子模型 膜使用高分辨率结构和冷冻密度，以及ENV的战略突变和截断 CT和MA。在AIM 3中，我们将利用保守的结构特征来设计由 膜可渗透的环状肽抑制剂。这些目标得到了全面的生化和 病毒学方法，将使用良好特征的进化枝C传播/创建者（T/F）HIV-1感染性 分子克隆完全代表了全局分离株，并呈现了完全匹配的Env和GAG 组成部分，可以区分我们的工作。我们的研究将以前所未有的细节揭示Env CT，MA和 膜脂质相互接合，在几乎本地情况下，所有组件都具有所有成分。这些强大的方法 将产生一个集成模型，包括难以解决的灵活区域，以实现完整的环境 Ma-Membrane机械栩栩如生。