Identifying determinants of HIV-1 responsible for the nanoscale distribution and dynamics of virus assembly

项目摘要 人类免疫缺陷病毒1型组装的详细机制 构成了我们对这种毁灭性的感染循环的认识上的重大差距 病原体。具体地说，基本包膜(Env)糖蛋白和 结构Gag分子有效结合形成具有感染性的HIV-1颗粒效果不佳 据了解，由于我们无法定量测量宇宙的时空动态 在受感染的宿主细胞内，单个分子处于纳米级。这项提案旨在划定 HIV-1在单分子基础上和在天然细胞环境中组装的基础。 为了实现这些目标，我们建议使用超分辨率显微镜来可视化和 量化病毒组装的各个阶段，以确定所发现的关键多肽基序的作用 在HIV-1Env和Gag中。我们的初步数据和分析表明，我们有能力 定量绘制HIV-1的生物发生图，并强烈表明这种方法将提供 对粒子组装的机制有明确的见解。这些发现将为我们提供新的 用于指导旨在挫败HIV-1组装的方法的量化模型。

Project Summary The detailed mechanisms of Human Immunodeficiency Virus Type 1 (HIV-1) assembly constitute a significant gap in our understanding of the infection cycle of this devastating pathogen. Specifically, the mechanisms by which the essential Envelope (Env) glycoprotein and structural Gag molecule efficiently coalesce to form an infectious HIV-1 particle are poorly understood, due to our inability to quantitatively measure the spatiotemporal dynamics of individual molecules at the nanoscale within infected host cells. This proposal aims to delineate the underpinnings of HIV-1 assembly on a single molecule basis and in a native cellular context. To accomplish these aims, we propose to utilize superresolution microscopy to visualize and quantify the stages of virus assembly in order to determine the role of key peptide motifs found in HIV-1 Env and Gag. Our preliminary data and analytics demonstrate our ability to quantitatively map the biogenesis of HIV-1 and strongly suggest that this approach will provide unequivocal insight in the mechanisms of particle assembly. These discoveries will provide new quantitative models to guide approaches aimed at thwarting HIV-1 assembly.