Retroviral Gag Trafficking, Env Incorporation, and Virus Assembly

逆转录病毒堵嘴贩运、Env 合并和病毒组装

• 批准号: 7733213
• 负责人: eric freed
• 金额: $ 46.06万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7733213
• 关键词: Adaptor Signaling Protein; Address; Cell membrane; Cell surface; Cells; Cytoplasmic Tail; Dependence; Destinations; Equine Infectious Anemia; Equine Infectious Anemia Virus; Equine Lentiviruses; Family; Family Felidae; Feline Immunodeficiency Virus; Gagging; Glycoproteins; Goals; HIV Drug Resistance Program; HIV-1; Imaging Techniques; Immunologic Deficiency Syndromes; Life; Lipids; Membrane Microdomains; Molecular; Phosphatidylinositols; Play; Point Mutation; Process; Reporting; Role; Site; Site Visit; Subfamily lentivirinae; Viral; Virion; Virus; Virus Assembly; base; cell type; cellular imaging; member; nonhuman primate; trafficking

In most cell types, HIV-1 assembly occurs predominantly on the plasma membrane; however, the itinerary that the Gag precursor follows to reach its destination in the cell remains ill-defined. It has been suggested that even in cell types in which assembly and budding occur predominantly at the plasma membrane, Gag traffics through an endosomal or endocytic compartment before reaching the cell surface. We and others have demonstrated that the matrix (MA) domain of Gag regulates the site of virus assembly, and we discovered recently that a member of the phosphoinositide family of lipids, phosphatidylinositol (4,5) bisphosphate [PI(4,5)P2], serves an important function in directing Pr55Gag to the plasma membrane. We are actively engaged in studies that seek to define the subcellular site of HIV-1 assembly in physiologically relevant cell types, and to elucidate further the cellular machinery involved in HIV-1 Gag trafficking. This effort makes use of live-cell imaging techniques recently developed in the VCIS, and is also being extended to the non-human primate lentiviruses equine infectious anemia virus (EIAV) and feline immunodeficiency virus (FIV). The viral envelope (Env) glycoproteins are incorporated into virions during the assembly process. We have shown that point mutations in the MA domain of Gag block Env incorporation, and demonstrated that the long gp41 cytoplasmic tail plays a cell type-dependent role in Env incorporation. Despite the evidence for a direct interaction between MA and the gp41 cytoplasmic tail, the molecular mechanism by which Env is incorporated, the subcellular site of initial Gag-Env contact, and the basis for cell type dependence of cytoplasmic tail function remain to be defined. We will address these issues; specifically, we will evaluate the role of cellular factors and lipid rafts in Env incorporation. [Corresponds to Freed Project 1 in the April 2007 site visit report of the HIV Drug Resistance Program]

在大多数细胞类型中，HIV-1的组装主要发生在质膜上；然而，Gag前体在细胞内到达目的地的路线仍然不明确。有研究表明，即使在组装和出芽主要发生在质膜上的细胞类型中，Gag在到达细胞表面之前也会通过内体或内胞室进行运输。我们和其他人已经证明Gag的基质（MA）结构域调节病毒组装位点，并且我们最近发现磷脂肌醇脂类家族的成员磷脂酰肌醇（4,5）二磷酸[PI(4,5)P2]在将Pr55Gag引导到质膜中起重要作用。我们正积极参与研究，试图在生理相关的细胞类型中定义HIV-1组装的亚细胞位点，并进一步阐明参与HIV-1 Gag运输的细胞机制。这项工作利用了VCIS最近开发的活细胞成像技术，并正在扩展到非人灵长类慢病毒马传染性贫血病毒（EIAV）和猫免疫缺陷病毒（FIV）。病毒包膜（Env）糖蛋白在组装过程中被整合到病毒粒子中。我们已经证明Gag的MA结构域的点突变阻断Env的结合，并证明长gp41细胞质尾部在Env的结合中起细胞类型依赖的作用。尽管有证据表明MA与gp41细胞质尾部之间存在直接相互作用，但Env结合的分子机制、Gag-Env初始接触的亚细胞位点以及细胞质尾部功能的细胞类型依赖性的基础仍有待明确。我们将解决这些问题；具体来说，我们将评估细胞因子和脂筏在Env掺入中的作用。[对应于2007年4月艾滋病毒耐药性计划实地考察报告中的自由项目1]

项目成果

Photoinduced reactivity of the HIV-1 envelope glycoprotein with a membrane-embedded probe reveals insertion of portions of the HIV-1 Gp41 cytoplasmic tail into the viral membrane.

HIV-1 包膜糖蛋白与膜嵌入探针的光诱导反应揭示了 HIV-1 Gp41 胞质尾部的部分插入到病毒膜中。

• DOI: 10.1021/bi701920f
• 发表时间: 2008
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Viard,Mathias;Ablan,SherimayD;Zhou,Ming;Veenstra,TimothyD;Freed,EricO;Raviv,Yossef;Blumenthal,Robert
• 通讯作者: Blumenthal,Robert