The Role of Rab6A in HIV-1 Env-mediated Entry into Monocyte-Derived Macrophages

Rab6A 在 HIV-1 Env 介导的进入单核细胞衍生巨噬细胞中的作用

• 批准号: 8732394
• 负责人: Isaac Zentner
• 金额: $ 4.12万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-27 至 2016-01-26
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8732394
• 关键词: Acquired Immunodeficiency Syndrome; Address; Adverse effects; Amino Acids; Architecture; Binding; Biological; Biological Assay; CCR5 gene; CD4 Antigens; Cancerous; Cell Line; Cell membrane; Cell physiology; Cells; Data; Dependency; Dominant-Negative Mutation; Drug Targeting; Early Endosome; Endoplasmic Reticulum; Event; GTP Binding; Genes; Genome; Goals; Golgi Apparatus; Guanine Nucleotide Exchange Factors; Guanosine Triphosphate Phosphohydrolases; HIV-1; Human; Infection; Integration Host Factors; Investigation; Lead; Life Cycle Stages; Macrophage Activation; Maintenance; Mediating; Monitor; Monomeric GTP-Binding Proteins; Nature; Pathogenesis; Pathway interactions; Physiological; Play; Process; Protein Isoforms; Proteins; Research; Role; Secretory Vesicles; Small Interfering RNA; Staging; Surface; Transport Process; Viral; Virus; base; cell type; combat; genome-wide; interest; macrophage; monocyte; mutant; novel; novel therapeutics; protein activation; protein function; public health relevance; rab GTP-Binding Proteins; receptor; research study; success; trafficking

DESCRIPTION (provided by applicant): The HIV-1 genome consists of only 9 genes and thus, depends highly on host cellular factors to replicate. Recent studies, through genome-wide siRNA screens, have identified many possible "HIV-1 dependency factors" HDFs. However, there lies a large inconsistency between these analyses and the majority of potential hits have not been verified in primary human target cells. Therefore, the goal of this proposal is to validat a recently identified possible HIV-1 Env-mediated host factor, Rab6A GTPase, in primary human monocyte-derived macrophages (MDMs) and define the mechanism by which this protein facilitates HIV-1 infection. Additionally, because Rab6A is ubiquitously expressed in two closely related isoforms, Rab6A and Rab6A', that have similar but distinct roles in the cell, both isoforms will be individually investigated. Similar to all GTPases, Rab6A(A/A') function is dependent on specific accessory proteins that allow cycling between active-GTP bound and inactive-GDP bound forms. We hypothesize that activated Rab6A(A/A') is required for efficient HIV-1 Env-mediated infection of MDMs and activation of Rab6A(A/A') is dependent on Ric1/Rgp1, a Rab6A(A/A')-specific guanine nucleotide-exchange factor, activity. We will utilize a pseudoviral single-round infection assay to define the mechanism by which Rab6A and Rab6A' facilitate HIV-1 replication in primary human macrophages. Our studies may reveal a novel therapeutic pathway to combat HIV-1/AIDS, as well as help dissect the fundamental biological role of Rab6A(A/A') in primary human macrophages.

描述（申请人提供）：HIV-1基因组仅由9个基因组成，因此高度依赖于宿主细胞因子进行复制。最近的研究，通过全基因组siRNA筛选，已经确定了许多可能的“HIV-1依赖因子”HDFs。然而，这些分析之间存在很大的不一致性，并且大多数潜在的命中尚未在原代人靶细胞中得到验证。因此，本提案的目标是验证最近在原代人单核细胞衍生的巨噬细胞（MDM）中鉴定的可能的HIV-1 Env介导的宿主因子Rab 6A GTdR，并确定该蛋白促进HIV-1感染的机制。此外，由于Rab 6A在两种密切相关的同种型Rab 6A和Rab 6A '中普遍表达，这两种同种型在细胞中具有相似但不同的作用，因此将单独研究这两种同种型。与所有GTP酶类似，Rab 6A（A/A '）功能依赖于允许在活性GTP结合形式和非活性GDP结合形式之间循环的特定辅助蛋白。我们假设激活的Rab 6A（A/A '）是HIV-1 Env介导的MDM有效感染所必需的，Rab 6A（A/A'）的激活依赖于Ric 1/Rgp 1（Rab 6A（A/A '）特异性鸟嘌呤核苷酸交换因子）活性。我们将利用假病毒单轮感染测定来确定Rab 6A和Rab 6A '促进HIV-1在原代人巨噬细胞中复制的机制。我们的研究可能揭示一种新的治疗途径来对抗HIV-1/AIDS，以及帮助剖析Rab 6A（A/A '）在原代人类巨噬细胞中的基本生物学作用。