HIV-1 INTEGRATION PROTEIN

HIV-1 整合蛋白

• 批准号: 2672061
• 负责人: DUANE P GRANDGENETT
• 金额: $ 19.84万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-01 至 2001-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2672061
• 关键词: electron microscopy; human immunodeficiency virus 1; human immunodeficiency virus 2; immunoprecipitation; integrase; recombinant DNA; virus infection mechanism

Integration of the human immunodeficiency virus type-1 (HIV-1) linear DNA genome into the host chromosome is essential for virus replication. The concerted insertion of the two viral DNA termini into the host genome (full-site reaction) requires the viral integrase (IN). Nonionic detergent lysates of viable HIV-1 virions can readily perform the in vivo full-site reaction using retrovirus-like donor substrates (469 bp) and circular DNA as target. Recombinant IN, purified from bacteria, can perform Only the half-site strand transfer reaction (insertion of only one viral terminus per target molecule). What properties associated with HIV-1 virions allow native IN to catalyze the full-site reaction? We will investigate the molecular mechanisms involved in how HIV-1 IN in virions and purified core particles, or IN purified from virions, can catalyze the 3' cleavage and concerted integration reactions. We will determine if other viral or cellular proteins associated with purified virions act as potential cofactors, either as enhancers or inhibitors, in the assembly and catalysis of preintegration complexes capable of full-site reactions. Efforts will be extended to determine whether an oligomeric structure of dimeric IN is responsible for catalyzing the concerted reaction. We will determine what role the viral terminal sequences have in the assembly of preintegration complexes, using native HIV-1 IN, that prompt the concerted reaction. Using DNA sequence analysis of donor-target recombinants which are genetically selected, we will investigate how native IN prompts the formation of full-site recombinants. We will investigate how an unknown recombinant structurally related to full-site recombinants and how specific sets of host site deletion (17-47 bp) recombinants, having a periodicity of approximately 10 bp between sets, are produced. Using electron microscopy, we will determine if native HIV-1 IN is capable of looping DNA which may play a role in forming and maintaining the 80S nucleoprotein complexes in vivo. Our studies may provide an understanding of the molecular mechanisms involved in the in vivo integration reaction and provide information for the design of IN inhibitors. Combinational inhibitor therapies against the rapid replication of HlV-1 in humans may be necessary to prevent AIDS.

人类免疫缺陷病毒1型（HIV-1）线性DNA的整合 将基因组插入宿主染色体对病毒复制至关重要。的 两个病毒DNA末端协同插入宿主基因组 （全位点反应）需要病毒整合酶（IN）。非离子洗涤剂 活HIV-1病毒体的裂解物可以容易地进行体内全位点 使用逆转录病毒样供体底物（469 bp）和环状DNA的反应 作为目标。从细菌中纯化的重组IN只能执行 半位点链转移反应（仅插入一个病毒末端 每个靶分子）。与HIV-1病毒体相关的哪些特性 天然IN来催化全位点反应我们将调查 HIV-1如何进入病毒体和纯化核心的分子机制 颗粒或从病毒体纯化的IN可以催化3'切割， 协调一致的一体化反应。我们将确定是否有其他病毒或 与纯化的病毒体相关的细胞蛋白作为潜在的 辅因子，作为增强剂或抑制剂，在组装中， 能够进行全位点反应的预整合复合物的催化。 将努力扩大，以确定是否低聚结构的 二聚IN负责催化协同反应。我们将 确定病毒末端序列在组装中的作用 整合前复合物，使用天然HIV-1 IN，促使协调的 反应利用供体-靶重组体的DNA序列分析， 是遗传选择，我们将调查如何在本地提示 形成全位点重组体。我们将调查一个未知的 重组体结构上与全位点重组体相关， 特定的宿主位点缺失（17-47 bp）重组体组，其具有 产生组之间约10 bp的周期性。使用 通过电子显微镜，我们将确定天然HIV-1 IN是否能够 环状DNA可能在形成和维持80 S 核蛋白复合物。我们的研究可能会提供一种理解 参与体内整合反应的分子机制 为IN抑制剂的设计提供了参考。组合 针对人类中HIV-1快速复制的抑制剂疗法可 是预防艾滋病的必要条件。