STRUCTURE/FUNCTION ANALYSIS OF THE HIV ENV GENE PRODUCT

HIV ENV 基因产物的结构/功能分析

• 批准号: 8172360
• 负责人: Eric Hunter
• 金额: $ 5.48万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8172360
• 关键词: Amino Acids; Area; Arginine; Biological Process; C-terminal; Cell fusion; Cell membrane; Cells; Charge; Complex; Computer Retrieval of Information on Scientific Projects Database; Cytoplasmic Tail; Funding; Glycine; Glycoproteins; Grant; HIV; Head; Human; Institution; Intracellular Transport; Investigation; Lipids; Lysine; Mediating; Membrane; Membrane Fusion; Modeling; Play; Proteins; Research; Research Personnel; Resources; Role; SIV; Series; Side; Source; Structure; Terminator Codon; Tryptophan; United States National Institutes of Health; Viral; Viral Pathogenesis; Virion; Virus; Virus Assembly; Virus Diseases; base; env Gene Products; mutant

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Three broad areas of investigation have been pursued: + Characterizing the role of the tryptophan-rich membrane-proximal (TRMP) domain of gp41 in Env-mediated membrane fusion and glycoprotein incorporation into virus. + Analyzing the topology and structural requirements of the gp41 membrane-spanning domain for virus assembly and entry. + Determining the role of sequences within the cytoplasmic domain (CD) of HIV Env in intracellular transport, and viral pathogenesis. The membrane-spanning domain (MSD) of the envelope (Env) glycoprotein from human (HIV) and simian immunodeficiency viruses plays a key role in anchoring the Env complex into the viral membrane but also contributes to its biological function in fusion and virus entry. In HIV type 1 (HIV-1), it has been predicted to span 27 amino acids, from lysine residue 681 to arginine 707, and encompasses an internal arginine at residue 694. By examining a series of C-terminal-truncation mutants of the HIV-1 gp41 glycoprotein that substituted termination codons for amino acids 682 to 708, we have shown that this entire region is required for efficient viral infection of target cells. Truncation to the arginine at residue 694 resulted in an Env complex that was secreted from the cells. In contrast, a region from residues 681 to 698, which contains highly conserved hydrophobic residues and glycine motifs and extends 4 amino acids beyond 694R, can effectively anchor the protein in the membrane, allow efficient transport to the plasma membrane, and mediate wild-type levels of cell-cell fusion. However, these fusogenic truncated Env mutants are inefficiently incorporated into budding virions. Based on the analysis of these mutants, a "snorkeling" model, in which the flanking charged amino acid residues at 681 and 694 are buried in the lipid while their side chains interact with polar head groups, is proposed for the HIV-1 MSD.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 已开展了三个广泛的调查领域： + 表征 gp41 富含色氨酸的近膜 (TRMP) 结构域在 Env 介导的膜融合和糖蛋白掺入病毒中的作用。 + 分析病毒组装和进入的 gp41 跨膜结构域的拓扑和结构要求。 + 确定 HIV Env 细胞质结构域 (CD) 内的序列在细胞内运输和病毒发病机制中的作用。 人类 (HIV) 和猿猴免疫缺陷病毒的包膜 (Env) 糖蛋白的跨膜结构域 (MSD) 在将 Env 复合物锚定到病毒膜中方面发挥着关键作用，但也有助于其在融合和病毒进入中的生物学功能。在 HIV 1 型 (HIV-1) 中，预计它跨越 27 个氨基酸，从赖氨酸残基 681 到精氨酸 707，并在残基 694 处包含一个内部精氨酸。 HIV-1 gp41 糖蛋白将终止密码子替换为氨基酸 682 至 708，我们已经证明这整个区域是有效病毒传播所必需的靶细胞的感染。在残基 694 处截断精氨酸会产生细胞分泌的 Env 复合物。 相比之下，残基 681 至 698 的区域包含高度保守的疏水残基和甘氨酸基序，并在 694R 之外延伸 4 个氨基酸，可以有效地将蛋白质锚定在膜上，允许有效转运到质膜，并介导野生型细胞与细胞融合的水平。然而，这些融合截短的 Env 突变体无法有效地整合到出芽的病毒体中。 基于对这些突变体的分析，提出了 HIV-1 MSD 的“浮潜”模型，其中 681 和 694 处的侧翼带电氨基酸残基埋在脂质中，而它们的侧链与极性头基相互作用。