Project 2. Immune evasion, trafficking, and nuclear import

项目 2. 免疫逃避、贩运和核进口

基本信息

批准号：
10653266
负责人：
Zandrea Ambrose
金额：
$ 86.22万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
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/10653266
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 Cytoplasm DNA Data Dependence Development Dynein ATPase Enzymes Genome Goals Guanine Nucleotide Exchange Factors HIV HIV-1 Immune Immune Evasion Immunologic Factors Infection Integration Host Factors Kinesin Knowledge Macrophage Mediating Methods Microtubule-Associated Proteins Microtubules Motor Mutation Mutation Analysis Nuclear Nuclear Import Nuclear Pore Complex Nuclear Pore Complex Proteins Pattern Process Proteins RNA Research Research Personnel Resolution Reverse Transcription Structure T-Lymphocyte Therapeutic Travel Tubulin Viral Viral Genome Viral Packaging Viral Proteins Virus Virus Diseases X-Ray Crystallography cell motility clinically relevant cofactor dynactin inhibitor integration site live cell microscopy molecular dynamics novel strategies particle protein transport recruit spatiotemporal superresolution imaging trafficking virology

项目摘要

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衣壳在病毒进入细胞核期间充当病毒与细胞之间的主要界面衣壳在复制周期的所有步骤中都至关重要，包括脱壳、宿主因素识别、微管运输、核输入和基因组整合。研究表明，HIV-1 衣壳劫持微管马达动力蛋白和驱动蛋白，使其朝着细胞核，完整的衣壳可以穿过细胞核孔复合体（NPC），这些数据显着。影响我们对 HIV-1 细胞质转运、核输入、核内转运和脱壳的看法，以及表明衣壳是 HIV-1 进入的关键参与者。然而，衣壳识别的原子水平理解。缺乏宿主因素的影响，并且尚不清楚病毒运动过程中因素的动态交换是如何发生的该项目的总体目标是填充这些从细胞外围到细胞核内的整合位点。通过提供有关衣壳参与免疫的关键结构和动态信息来弥补知识差距拟议的研究是七个 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），具体来说，我们将 i)。阐明 HIV-1 衣壳与宿主先天免疫蛋白（包括 MxB 和 TRIM5α/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/dynactin/BICD2，以及微管；以及 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相互作用，包括核孔蛋白 Nup358 和Nup153以及宿主因子CPSF6和SUN1/2我们的结果将提供前所未有的原子水平。衣壳与宿主蛋白相互作用的结构和动态视图，可以指导新的开发临床相关的衣壳抑制剂。