Dynamics of Gag-Pol auto-processing and ESCRT recruitment during HIV budding

项目总结： 我们的长期目标是完全了解调节释放的分子机制 从被感染的细胞中分离出具有传染性的HIV病毒粒子。在我们之前的资助期(R21)中，我们发现早期 运输所需的内体分选复合体在病毒粒子之间的分子竞赛中发挥着重要作用 萌芽和Gag-Pol自动处理。我们还发现，干扰萌芽和自动之间的平衡- 处理会导致非传染性病毒粒子的释放。面对蛋白水解酶作用下HIV的快速进化 对于抑制剂，寻找新的靶向机制以遏制感染至关重要。GAG-POL自动处理及其实现 组装和萌芽期间的监管是一个耐人寻味的目标。我们对此的基本理解 然而，由于其与组装和萌芽动力学的纠缠以及不同步，工艺受到限制 病毒粒子释放的性质。在这里，我们建议可视化HIV萌芽和Gag-Pol自动处理在单个 体内的病毒粒子：1)建立完整HIV病毒粒子期间ESCRT早期募集的动态和调节 组装，2)测量HIV萌发过程中Gag-Pol掺入和自动处理的动态，3) 剖析了GAG-POL自动加工的规则。

Project Summary: Our long-term goal is to gain a complete understanding of the molecular machinery that regulates the release of infectious HIV virions from infected cells. During our previous funding period (R21), we found that early Endosomal Sorting Complexes Required for Transport play a major role in a molecular race between virion budding and Gag-Pol auto-processing. We also found that disturbing the balance between budding and auto- processing results in release of non-infectious virions. In the face of rapid evolution of HIV under protease inhibitors, search for new target mechanisms to curb infections is crucial. Gag-Pol auto-processing and its regulation during assembly and budding present an intriguing target. Our fundamental understanding of this process however is limited due to its entanglement with assembly and budding kinetics and asynchronous nature of virion release. Here we propose to visualize HIV budding and Gag-Pol auto-processing in single virions in vivo to: 1) Establish the dynamics and regulation of early ESCRT recruitment during full HIV virion assembly, 2) Measure the dynamics of Gag-Pol incorporation and auto-processing during HIV budding and 3) Dissect the regulation of Gag-Pol auto-processing.