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组装主要发生在质膜上。 但是，插口前体遵循的行程仍未定义。已经提出，即使在质膜中主要发生组装和萌芽的细胞类型中，在到达细胞表面之前，通过内体或内吞室进行了堵嘴。我们和其他人已经证明，GAG的矩阵（MA）结构域调节病毒组装的部位，最近我们发现，脂质磷酸固醇家族的成员，磷脂酰肌醇（4,5）双磷酸[PI（4,5）P2]在PR55GAG中起着重要功能。我们积极从事试图在生理相关的细胞类型中定义HIV-1组装的亚细胞位点的研究，并进一步阐明与HIV-1 GAG运输相关的细胞机械。这项工作利用了最近在VCIS开发的活细胞成像技术，并且也扩展到非人类灵长类动物慢病毒，马匹传染病病毒（EIAV）和猫免疫缺陷病毒（FIV）。 在组装过程中，病毒包膜（Env）糖蛋白被掺入病毒粒子中。我们已经表明，在GAG块环境的MA域中，点突变，并证明长GP41细胞质尾巴在Envimation中起着细胞类型依赖性作用。尽管有证据表明MA和GP41细胞质尾巴之间存在直接相互作用，但结合了ENV的分子机制，初始GAG-ENV接触的亚细胞位点，以及细胞类型依赖性细胞质尾部功能的基础。我们将解决这些问题；具体而言，我们将评估细胞因子和脂质筏在Envimation中的作用。 [对应于2007年4月的艾滋病毒抗药性计划的现场访问报告中的释放项目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