HIV Release and Restriction

HIV 释放和限制

• 批准号: 10221479
• 负责人: David Bulkley
• 金额: $ 78.58万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-27 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10221479
• 关键词: ATP phosphohydrolase; Antiviral Agents; Binding; Biochemical; Carrier Proteins; Cells; Cellular biology; Complex; Crowding; Cryoelectron Microscopy; Cytoskeletal Filaments; Electrons; Elements; Event; Face; Family member; Filament; Fluorescence; Genes; HIV; HIV Budding; HIV-1; Human; Image; Individual; Integration Host Factors; Letters; Lipids; Membrane; Methods; Modeling; Mus; Nature; Neck; Pathway interactions; Peptides; Polymers; Primates; Process; Proteins; Resolution; Retrotransposition; Rodent; Saimiri; Site; Sorting - Cell Movement; Structure; Testing; Total Internal Reflection Fluorescent; Toxic effect; Variant; Viral; Virion; Virus Replication; Yeasts; base; blocking factor; cofactor; constriction; design; light microscopy; membrane model; mouse genome; novel; reconstruction; recruit; scaffold; structural biology; trafficking; virus envelope

ABSTRACT This Project is designed to elucidate the mechanistic bases for HIV-1 budding and its inhibition by the host factor retroCHMP3. To initiate budding, HIV-1 Gag binds ESCRT-I1-4 and ALIX5-7, two early-acting factors in the host Endosomal Sorting Complexes Required for Transport (ESCRT) pathway. These factors, in turn, recruit the late-acting ESCRT factors CHMP2, CHMP4 (ESCRT-III) and VPS4 (AAA ATPase)2,8-10 (reviewed in refs. 11-16). Numerous models for membrane constriction and fission have been proposed.17-26 We favor those in which: 1) ESCRT-III proteins form spiraling filaments that recruit VPS4 and constrict the membrane neck from the cytoplasmic face and 2) The VPS4 ATPase promotes lipid mixing by remodeling the underlying ESCRT-III scaffold. We now propose to determine the structural bases for membrane remodeling by ESCRT-III filaments and VPS4 (Aims 1 and 2), characterize how mammalian retroCHMP3 (ESCRT-III) proteins can inhibit HIV budding without inducing cellular toxicity (Aim 3), and image ESCRT assemblies within budding virions by electron cryotomography (Aim 4). These studies will reveal how HIV-1 uses the host ESCRT-III/VPS4 machinery to exit cells and how host retroCHMP3 proteins can specifically inhibit this process.

抽象的 该项目旨在阐明 HIV-1 出芽及其被宿主抑制的机制基础 因子retroCHMP3。为了启动出芽，HIV-1 Gag 结合 ESCRT-I1-4 和 ALIX5-7，这两个早期作用因子 运输所需的宿主内体分选复合物（ESCRT）途径。这些因素反过来又招募 晚效 ESCRT 因子 CHMP2、CHMP4 (ESCRT-III) 和 VPS4 (AAA ATPase)2,8-10（参考文献中综述） 11-16）。人们已经提出了许多膜收缩和裂变的模型。17-26 我们赞成以下领域的模型： 其中：1) ESCRT-III 蛋白形成螺旋丝，招募 VPS4 并收缩膜颈 细胞质面和 2) VPS4 ATPase 通过重塑潜在的 ESCRT-III 促进脂质混合 脚手架。我们现在建议确定 ESCRT-III 丝进行膜重塑的结构基础 和VPS4（目标1和2），描述哺乳动物retroCHMP3 (ESCRT-III)蛋白如何抑制HIV 出芽而不诱导细胞毒性（目标 3），并通过以下方法对出芽病毒粒子内的 ESCRT 组件进行成像 电子冷冻断层扫描（目标 4）。这些研究将揭示 HIV-1 如何利用宿主 ESCRT-III/VPS4 退出细胞的机制以及宿主retroCHMP3蛋白如何特异性抑制这一过程。