Project 2. Immune evasion, trafficking, and nuclear import

项目 2. 免疫逃避、贩运和核进口

• 批准号: 10506953
• 负责人: Zandrea Ambrose
• 金额: $ 74.2万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10506953
• 关键词: Amino Acid Substitution; Anti-Retroviral Agents; Binding; Biochemical; Biological Assay; CD4 Positive T Lymphocytes; Capsid; Capsid Proteins; Cell Nucleus; Cell membrane; Cells; Complement; Complex; Cryoelectron Microscopy; Cyclophilin A; DNA; Data; Dependence; Development; Dynein ATPase; Enzymes; Genome; Goals; Guanine Nucleotide Exchange Factors; HIV; HIV-1; Image; Immune; Immune Evasion; Immunologic Factors; Infection; Integration Host Factors; Kinesin; Knowledge; Mediating; Methods; Microtubule-Associated Proteins; Microtubules; Motor; Mutation; Mutation Analysis; Nuclear; Nuclear Import; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Pattern; Process; Protein Dynamics; Proteins; RNA; Research; Research Personnel; Resolution; Reverse Transcription; Structure; T-Lymphocyte; TRIM Gene; Therapeutic; Travel; Tubulin; Viral; Viral Genome; Viral Proteins; Virus; Virus Diseases; X-Ray Crystallography; cell motility; clinically relevant; dynactin; inhibitor; integration site; live cell microscopy; macrophage; molecular dynamics; novel strategies; particle; protein transport; recruit; spatiotemporal; trafficking; virology

Project 2 – Immune evasion, trafficking and nuclear import The HIV-1 capsid acts as the primary interface between the virus and the cell during viral ingress and nuclear entry. The capsid is critically involved throughout all steps of the replication cycle, including uncoating, recognition by host factors, trafficking along microtubules, nuclear import, and genome integration. Recent research revealed that HIV-1 capsid hijacks microtubule motors dynein and kinesin for its journey towards the nucleus, and an intact capsid can traverse the cellular nuclear pore complex (NPC). These data significantly impact our views of HIV-1 cytoplasmic transport, nuclear import, intranuclear transport, and uncoating, and indicate that capsid is a key player in HIV-1 ingress. However, atomic-level understanding of capsid recognition by host factors is lacking, and it is unclear how the dynamic exchange of factors occurs during viral movement from the cell periphery to the site of integration inside the nucleus. The overall goal of this project is to fill these knowledge gaps by providing critical structural and dynamic information on capsid’s engagement in immune evasion, trafficking and integration. The proposed studies are an integrated effort from seven PCHPI investigators and four cores (CryoEM/ET, NMR, Computational, and HIV Virology). Specifically, we will i) elucidate the interactions of HIV-1 capsid with host innate immune proteins, including MxB and TRIM5α/TRIMCyp; ii) determine the interactions between HIV-1 capsid and intracellular trafficking proteins, including kinesin-1/FEZ1 and dynein/dynactin/BICD2, and microtubules; and iii) define the dynamics of HIV-1 capsid interactions with host dependency factors involved in nuclear import, including the nucleoporins Nup358 and Nup153 and the host factors CPSF6 and SUN1/2. Our results will provide unprecedented atomic-level structural and dynamic view of capsid’s interactions with host proteins and could guide the development of new clinically relevant capsid inhibitors.

项目2 -免疫逃避、贩运和核进口 HIV-1衣壳在病毒侵入和核扩散期间充当病毒和细胞之间的主要界面。 入境衣壳在整个复制周期的所有步骤中起关键作用，包括脱壳， 宿主因子的识别、沿沿着微管的运输、核输入和基因组整合。最近 研究表明，HIV-1衣壳劫持微管马达动力蛋白和驱动蛋白， 完整的衣壳可以穿过细胞核孔复合体（NPC）。这些数据显著 影响我们对HIV-1胞质转运、核输入、核内转运和脱壳的看法， 表明衣壳在HIV-1侵入中起关键作用。然而，对衣壳识别的原子级理解 缺乏宿主因子的影响，并且不清楚在病毒运动期间因子的动态交换如何发生 从细胞外围到细胞核内的整合位点。该项目的总体目标是填补这些 通过提供关于衣壳参与免疫反应的关键结构和动态信息， 逃避、贩运和融合。拟议的研究是七个PCHPI的综合努力 研究人员和四个核心（CryoEM/ET，NMR，计算和HIV病毒学）。具体而言，我们将（i） 阐明HIV-1衣壳与宿主先天免疫蛋白（包括MxB和 TRIM 5 α/TRIMCyp; ii）确定HIV-1衣壳和细胞内运输蛋白之间的相互作用， 包括驱动蛋白-1/FEZ1和动力蛋白/动力肌动蛋白/BICD 2，和微管;和iii）定义HIV-1的动力学 衣壳与参与核输入的宿主依赖性因子的相互作用，包括核孔蛋白Nup 358 和NUP 153以及宿主因子CPSF 6和SUN 1/2。我们的研究结果将提供前所未有的原子级 衣壳与宿主蛋白相互作用的结构和动态视图，并可以指导新的 临床相关的衣壳抑制剂。