HIV Release and Restriction

HIV 释放和限制

• 批准号: 9411508
• 负责人: Adam Frost
• 金额: $ 65.19万
• 依托单位: UNIVERSITY OF UTAH
• 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/9411508
• 关键词: ATP phosphohydrolase; Antiviral Agents; Binding; Biochemical; Carrier Proteins; Cells; Cellular biology; Complex; Crowding; 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; Recruitment Activity; 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; protein transport; reconstruction; 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发芽的机械基础及其抑制宿主的抑制作用 因子retrochmp3。为了发芽，HIV-1 GAG结合了ESCRT-I1-4和ALIX5-7，这是两个早期作用因素 运输（ESCRT）途径所需的宿主内体分类复合物。这些因素反过来招募 晚期作用的ESCRT因子CHMP2，CHMP4（ESCRT-III）和VPS4（AAA ATPase）2,8-10（在参考文献中进行了审查。 11-16）。已经提出了许多用于膜收缩和裂变的模型。17-26我们有利于那些 其中：1）ESCRT-III蛋白质形成螺旋丝，募集VPS4并从膜上收缩 细胞质面和2）VPS4 ATPase通过重塑下面的ESCRT-III促进脂质混合 脚手架。现在，我们建议确定通过ESCRT-III丝进行膜重塑的结构碱基 和VPS4（目标1和2），表征了哺乳动物retrochmp3（ESCRT-III）蛋白如何抑制HIV 萌芽而无需诱导细胞毒性（AIM 3），并通过 电子冷冻学（AIM 4）。这些研究将揭示HIV-1如何使用宿主ESCRT-III/VPS4 向退出细胞的机械以及宿主retrochmp3蛋白如何特异性抑制此过程。