Defining the nuclear import pathways of HIV-1

定义 HIV-1 的核输入途径

基本信息

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
项目类别：
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

项目摘要

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.

抽象的像所有灵长类动物一样，人类免疫缺陷病毒（HIV-1）具有感染能力通过与核孔复合物的成分接合并介导的细胞病毒核糖核蛋白复合蛋白（RNP）的核易位，随后整合进入宿主细胞基因组。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）的成分以完成核过程进口。不幸的是，HIV-1通过核孔易位的确切机制复杂仍然是病毒生命周期中最不理知识的步骤之一。此外，最近支持NPC的概念的证据比以前想象的更异质进一步混淆了这种情况。我们已经开发了一个可诱导的核孔封锁允许对任何功能性，传染性病毒基因组的核进口率进行监测相关细胞类型。使用此技术，我们观察到某些CA突变体对 NUP62介导的细胞中介导的核孔隙阻塞，该细胞有效地阻断了野生型CA的感染，证明HIV-1可以在感染过程中利用独特的核进口途径。在这个应用，我们将确定NPC的异质程度并绘制特定的程度核孔成分构成HIV-1在核进入过程中使用的NPC。与我们的核孔封锁，我们现在有能力使用不同的核孔阻止NPC 成分。因此，该应用旨在定义和可视化特定的核孔相互作用并介导HIV-1核进口的成分以及特定的NPC使用方式影响靶细胞中的病毒整合。总的来说，此应用程序将弥合与我们的关键差距了解HIV-1的核进口。