Diverse Functions of HIV-1 Capsid During Postentry Events

项目摘要 HIV-1衣壳包裹着病毒和宿主感染所必需的成分， 每个进入后步骤，包括逆转录、核进入和整合靶向。这些衣壳- 介导的事件由HIV-1脱壳控制，在这个过程中，锥形核心脱落病毒衣壳 蛋白质（CA），并转化为逆转录复合物，然后预整合复合物。 病毒衣壳的脱壳受核心稳定性和衣壳结合宿主辅因子的调节。多 如何协调去包被过程以促进病毒繁殖仍有待了解。时间 抗病毒药物引起的衣壳解体（延迟或加速脱壳）的正常过程偏离 因子或药理学操作可严重损害病毒感染性。然而，目前仍不清楚 核心稳定性如何影响衣壳解体和下游事件。我们的初步 研究已经确定了一种天然的HIV-1毒株，该毒株似乎改变了核心稳定性，而不损害病毒 健身在目标1中，我们将描述这种独特变体的广泛生物学特性， 利用靶向衣壳的抗病毒药物的核心去稳定活性作为研究衣壳稳定性的工具。现款车型 证实核心分解影响了HIV-1 DNA的先天感受，但机制细节尚未确定。 阐明。在目标2中，我们建议通过操纵核心稳定性和 衣壳与宿主蛋白的相互作用影响先天免疫应答。衣壳脱壳也在 核事件，如核进口和一体化目标。在目标3中，我们将讨论这些事件如何 在功能上相互关联，并通过使用各种实验方法促进最佳病毒生长， 包括新的高通量单细胞测定。

Project Summary The HIV-1 capsid, which encloses viral and host components essential for establishing infection, influences every post-entry step, including reverse transcription, nuclear entry and integration targeting. These capsid- mediated events are governed by HIV-1 uncoating, a process in which the conical core sheds viral capsid proteins (CA) and transforms into reverse transcription complexes and then pre-integration complexes. Uncoating of the viral capsid is regulated by both core stability and capsid-binding host cofactors. Much remains to be learned about how the uncoating process is coordinated to facilitate viral propagation. Temporal deviations from the normal course of capsid disassembly (delayed or accelerated uncoating) by antiviral factors or pharmacological manipulation can severely impair viral infectivity. However, it still remains unclear precisely how intrinsic core stability affects capsid disassembly and downstream events. Our preliminary studies have identified a natural HIV-1 strain that appears to alter core stability without compromising viral fitness. In Aim 1, we will characterize a broad range of biological properties of this unique variant and also exploit core-destabilizing activity of capsid-targeting antivirals as a tool to study capsid stability. Current models posit that core disassembly impacts innate sensing of HIV-1 DNA but mechanistic details have yet to be elucidated. In Aim 2, we propose to examine how perturbation of uncoating by manipulating core stability and capsid interactions with host proteins affects innate immune responses. Capsid uncoating also plays roles in nuclear events, such as nuclear import and integration targeting. In Aim 3, we will address how these events are functionally linked and contribute to optimal viral growth by using various experimental approaches, including novel high-throughput single-cell assays.