Defining the nuclear import pathways of HIV-1

摘要 人类免疫缺陷病毒(HIV-1)像所有灵长类慢病毒一样，具有感染 通过与核孔复合体的成分接触并介导细胞分裂而不分裂细胞 病毒核糖核蛋白复合体(RNP)的核移位以便随后整合 进入宿主细胞基因组。病毒衣壳蛋白(CA)与多种宿主因子相互作用 使核孔复合体(NPC)的成分参与完成核的过程 导入。不幸的是，HIV-1通过核孔转移的确切机制 复杂性仍然是病毒生命周期中最鲜为人知的步骤之一。此外，最近 支持NPC概念的证据比之前认为的更不同 这进一步扰乱了这种情况。我们已经开发出一种可诱导的核孔阻断 允许对功能性、传染性病毒基因组的核进口速度进行监测 相关单元格类型。使用这项技术，我们观察到某些CA突变体对 A Nup62介导的细胞核孔阻断可有效阻断野生型CA感染， 表明艾滋病毒-1可以在感染期间利用不同的核进口途径。在这 应用程序，我们将确定NPC的异构性程度，并将特定的 构成HIV-1在核进入过程中利用的NPC的核孔成分。带着我们的 核孔阻断，我们现在有能力利用不同的核孔来阻断NPC 选民。因此，该应用程序旨在定义和可视化特定的核孔 相互作用和调节HIV-1核进口的成分以及NPC的具体用途 影响病毒在靶细胞中的整合。总体而言，这一应用程序将填补与我们的 对核进口艾滋病毒-1的了解。

Abstract Human Immunodeficiency virus (HIV-1), like all primate lentiviruses possesses the ability to infect non-dividing cells by engaging with components of the nuclear pore complex and mediating the nuclear translocation of the viral ribonucleoprotein complex (RNP) for subsequently integration into the host cell genome. The viral capsid protein (CA) interacts with numerous host factors involving constituents of the nuclear pore complex (NPC) to accomplish the process of nuclear import. Unfortunately, the exact mechanism by which HIV-1 translocates through the nuclear pore complex remains one of the least understood steps of the viral life cycle. In addition, recent evidences supporting the notion of NPCs being more heterogeneous than previously thought further confounds this situation. We have developed an inducible nuclear pore blockade that allows the rate of nuclear import of functional, infectious viral genomes to be monitored in any relevant cells types. Using this technique, we observe that certain CA mutants are insensitive to a Nup62 mediated nuclear pore blockade in cells which potently block infection by wild type CA, demonstrating that HIV-1 can utilize distinct nuclear import pathways during infection. In this application, we will determine the degree to which NPC are heterogeneous and map the specific nuclear pore constituents that makeup the NPCs utilized by HIV-1 during nuclear entry. With our nuclear pore blockade, we now have the capability to block NPCs using different nuclear pore constituents. As such, this application aims to define and visualize the specific nuclear pore constituents that interact and mediate the nuclear import of HIV-1 and how specific NPC usage affects viral integration in target cells. Collectively this application would close critical gaps to our understanding in to the nuclear import of HIV-1.