HIV-1 INTEGRATION PROTEIN

人类免疫缺陷病毒1型(HIV-1)线性DNA的整合 基因组进入宿主染色体是病毒复制的关键。这个 将两个病毒DNA末端协同插入宿主基因组 (全部位反应)需要病毒整合酶(IN)。非离子洗涤剂 存活的HIV-1病毒粒子的裂解物可以很容易地在体内进行全部位 利用类逆转录病毒供体底物(469bp)和环状DNA的反应 作为目标。从细菌中提纯的重组IN只能执行 半位点链转移反应(仅插入一个病毒末端 每个目标分子)。与HIV-1病毒粒子相关的特性允许 天然IN能催化全位反应吗？我们将调查 HIV-1进入病毒粒子和纯化核心的分子机制 粒子，或从病毒粒子提纯的IN，可以催化3‘裂解和 协调一致的整合反应。我们将确定是否有其他病毒或 与纯化的病毒粒子相关的细胞蛋白是潜在的 辅因子，作为促进剂或抑制剂，在组装和 能够全位反应的前整合络合物的催化作用。 将继续努力确定一种寡聚体结构是否 二聚体IN负责催化协同反应。我们会 确定病毒末端序列在装配过程中起什么作用 整合前复合体，使用本地HIV-1 IN，促进协调 反应。利用DNA序列分析的供体-靶向重组体 是通过基因选择的，我们将调查原生IN是如何提示 形成全位点重组体。我们将调查一个未知的 重组在结构上与全部位重组体的关系以及如何 宿主位点缺失(17-47bp)重组体的特定集合，具有 在集合之间产生大约10BP的周期。vbl.使用 电子显微镜，我们将确定天然HIV-1IN是否有能力 可能在80年代的形成和维持中起作用的环状DNA 体内的核蛋白复合体。我们的研究可能会提供一种理解 体内整合反应涉及的分子机制 为IN抑制剂的设计提供参考。组合式 针对人类HLV-1快速复制的抑制剂疗法可能 对预防艾滋病来说是必要的。

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.