Defining the nuclear import pathways of HIV-1

定义 HIV-1 的核输入途径

• 批准号: 10641980
• 负责人: Edward M Campbell
• 金额: $ 72.84万
• 依托单位: LOYOLA UNIVERSITY CHICAGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-23 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10641980
• 关键词: Affect; Architecture; Area; Automobile Driving; Binding; Binding Proteins; Biochemical; Biological Assay; Biology; Capsid; Capsid Proteins; Cell Nucleus; Cells; Chromatin; Cleavage And Polyadenylation Specificity Factor; Complement; Complex; Cyclophilin A; Cytoplasm; Data; Dimerization; Electron Microscopy; Engineering; Environment; Event; Genes; Genetic Transcription; Genome; HIV; High-Throughput Nucleotide Sequencing; Histones; Image; Individual; Infection; Infection prevention; Integration Host Factors; Interphase Cell; Knowledge; Lentivirus; Life Cycle Stages; Light; Lighting; Maps; Mediating; Methodology; Microscopy; Monitor; Mutation; Nuclear; Nuclear Accidents; Nuclear Import; Nuclear Localization Signal; Nuclear Pore; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Nuclear Protein; Nuclear Translocation; Pathway interactions; Pharmaceutical Preparations; Population; Pore Proteins; Primate Lentiviruses; Process; Proviruses; Research; Reverse Transcription; Ribonucleoproteins; Role; Series; Staging; Structure; Techniques; Therapeutic Intervention; Viral; Viral Genome; Viral Physiology; Virus; Virus Diseases; Virus Integration; Visualization; cell type; experimental study; imaging approach; insight; integration site; mutant; novel; reactivation from latency; spatiotemporal; therapeutic development; trafficking

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.

抽象的 人类免疫缺陷病毒 (HIV-1) 与所有灵长类慢病毒一样具有感染能力 通过与核孔复合物的成分结合并介导非分裂细胞 病毒核糖核蛋白复合物 (RNP) 的核转位以便随后整合 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 涉及核孔复合体（NPC）的成分来完成核化过程 进口。不幸的是，HIV-1 通过核孔移位的确切机制 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 比之前想象的更加异质这一观点的证据 使这种情况更加混乱。我们开发了一种诱导性核孔阻断剂 允许在任何环境中监测功能性、传染性病毒基因组的核输入率 相关细胞类型。使用这种技术，我们观察到某些 CA 突变体对 Nup62 介导的细胞核孔阻断，可有效阻断野生型 CA 的感染， 证明 HIV-1 在感染过程中可以利用不同的核输入途径。在这个 应用程序中，我们将确定 NPC 的异质程度并绘制具体的图 构成 HIV-1 在进入核过程中利用的 NPC 的核孔成分。与我们的 核孔封锁，我们现在有能力使用不同的核孔封锁NPC 选民。因此，该应用程序旨在定义和可视化特定的核孔 相互作用并介导 HIV-1 核输入的成分以及 NPC 的具体用途 影响病毒在靶细胞中的整合。总的来说，这个应用程序将缩小与我们的关键差距 了解 HIV-1 的核输入。

项目成果

Teaching old dogmas new tricks: recent insights into the nuclear import of HIV-1.

教旧教条新技巧：对 HIV-1 核输入的最新见解。

• 发表时间: 2022-04
• 作者: Dharan, Adarsh;Campbell, Edward M
• 通讯作者: Campbell, Edward M