Evolutionary potential of HIV-1 capsid: mechanisms and consequences

项目总结 病毒衣壳是一种锥形的蛋白外壳，在HIV-1复制过程中扮演着多种角色，并已出现 作为一个很有前途的抗病毒靶点。人们普遍认为，结构和功能的限制限制了 衣壳的突变耐受性。然而，人们越来越认识到，艾滋病毒-1适应了 在不影响病毒复制适合性的情况下，几乎每个衣壳介导的功能都具有表型可变性。 这些观察结果表明，HIV-1衣壳的适应能力可能被低估了。 也很充实。HIV-1衣壳的进化潜力与耐药性和 作为寄主因子利用和逃避的分子基础。在此应用程序中，基于我们的 在之前和初步的研究中，我们建议研究HIV-1衣壳的机制和后果 采用多管齐下的战略，特别注重使用大规模遗传方法，以提高可变性。在……里面 目标1，将对HIV-1衣壳进行高通量突变分析，以产生全面的 不同衣壳蛋白抑制剂的耐药谱。我们还将探索不同类型的组合用法如何 衣壳蛋白抑制剂可以通过协同作用和/或通过限制 耐药途径。在目标2中，阐明HIV-1如何通过高度带电的毛孔利用多种宿主因素 在每个CA多聚体的中心，我们将研究这些细胞因子对衣壳稳定性的要求 以及不同慢病毒之间的其他进入后步骤。此外，深度突变扫描和详细的 将对HIV-1衣壳的中央毛孔进行表型鉴定。在目标3中，利用艾滋病毒-1的可获得性 变异体具有不同的脱壳动力学，我们将研究衣壳在保护病毒DNA免受天然感染方面的作用 在艾滋病毒-1细胞间传播的背景下的传感器。第二，确定经济衰退的影响 病毒DNA上的衣壳可以决定病毒感染的命运，我们将开发新的实验检测方法 这使得在共培养系统中能够量化病毒潜伏期和竞争。这些研究将会增加 我们对衣壳介导功能的分子基础的理解。新的发现将提供有价值的 对HIV-1衣壳进化潜力的洞察，并可能被利用来帮助发现和 新型靶向抗病毒药物衣壳的设计。

PROJECT SUMMARY The viral capsid, a cone-shaped protein shell, plays multiple roles during HIV-1 replication and has emerged as a promising antiviral target. It has been widely accepted that structural and functional constraints limit the mutational tolerance of the capsid. However, there is growing appreciation that HIV-1 accommodates phenotypic variability in nearly every capsid-mediated function without compromising viral replicative fitness. These observations raise the possibility that the capability of HIV-1 capsid to adapt is likely underestimated and substantial. The evolutionary potential of HIV-1 capsid has direct implications for drug resistance and serves as the molecular basis for host factor utilization and evasion. In this application, building upon our previous and preliminary studies, we propose to study the mechanisms and consequences of HIV-1 capsid variability using a multi-pronged strategy with special focus on the use of large-scale genetic approaches. In Aim 1, high-throughput mutational analysis of HIV-1 capsid will be conducted to generate comprehensive resistance profiles of various capsid inhibitors. We will also explore how combinatorial use of different types of capsid inhibitors can improve antiviral activity through synergistic interactions and/or by restricting resistance pathways. In Aim 2, to elucidate how HIV-1 exploits multiple host factors via highly charged pores at the center of each CA multimer, we will study the requirement for these cellular factors in capsid stability and other post-entry steps among diverse lentiviruses. Additionally, deep mutational scanning and detailed phenotyping will be applied to the central pore of HIV-1 capsid. In Aim 3, leveraging the availability of HIV-1 variants with varying uncoating kinetics, we will study the role of the capsid in shielding viral DNA from innate sensors in the context of cell-to-cell transmission of HIV-1. Second, to determine how the impacts of the capsid on viral DNA sensing can define the fate of viral infection, we will develop novel experimental assays that enable quantification of viral latency and competitions in a co-culture system. These studies will increase our understanding of the molecular basis for capsid-mediated functions. New findings will provide valuable insights into the evolutionary potential of HIV-1 capsid and may be harnessed to aid in the discovery and design of novel capsid targeting antivirals.