Defining the nuclear import pathways of HIV-1

摘要 人类免疫缺陷病毒（HIV-1），像所有的灵长类慢病毒一样，具有感染 通过与核孔复合物的组分接合并介导非分裂细胞， 病毒核糖核蛋白复合物（RNP）的核转位，随后整合 插入宿主细胞基因组中。病毒衣壳蛋白（CA）与许多宿主因子相互作用 涉及核孔复合物（NPC）的成分，以完成核的过程， 导入.不幸的是，HIV-1通过核孔转移的确切机制 复合体仍然是病毒生命周期中最不为人所知的步骤之一。此外，最近 支持NPC比以前认为的更异质的概念的证据 进一步混淆了这种情况。我们开发了一种可诱导的核孔阻断剂， 允许功能性、感染性病毒基因组的核输入速率在任何 相关细胞类型。使用这种技术，我们观察到某些CA突变体对 Nup 62介导的细胞核孔阻断，其有效阻断野生型CA的感染， 这表明HIV-1在感染过程中可以利用不同的核输入途径。在这 应用程序，我们将确定NPC异质性的程度，并映射特定的 构成HIV-1在进入核期间利用的NPC的核孔成分。与我们 核孔封锁，我们现在有能力阻止NPC使用不同的核孔 选民。因此，本申请旨在定义和可视化特定的核孔 相互作用和介导HIV-1的核输入的成分，以及特定的NPC使用 影响靶细胞中的病毒整合。总的来说，这一应用程序将缩小我们与 了解HIV-1的核输入。

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.