GENETICS AND MECHANISM OF ACTION OF THE HIV INTEGRASE

HIV 整合酶的遗传学和作用机制

• 批准号: 3146386
• 负责人: WILLIAM A. HASELTINE
• 金额: $ 18.37万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-02-01 至 1997-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3146386
• 关键词: DNA binding protein; SDS polyacrylamide gel electrophoresis; cinefluorography; circular DNA; human immunodeficiency virus 1; nucleic acid repetitive sequence; protein purification; southern blotting; tissue /cell culture; virus DNA; virus RNA; virus genetics; virus protein

The goal of this proposal is to obtain an understanding of the fate of viral DNA after the viral RNA has been converted to DNA by the concerted action of the viral DNA polymerase and ribonuclease activities. The two objectives of this proposal are to understand the process of integration of viral DNA into host DNA and to understand the process of formation of virus DNA circles. Our preliminary work demonstrates that both the integration and circularization reactions can be studied in vitro. Objective 1. Characterization of the integration reaction. Specific Aims include: A. Characterization of virus and cell specified proteins in the preintegration complexes that are present in the cytoplasm and the nucleus of the newly infected cell. B. Reconstruction of an efficient integration reaction using purified virus and cell proteins. Objective 2. Understanding the reactions which govern formation of viral DNA circles. Three types of viral DNA circles are made in infected cells, 1-LTR circles, simple 2-LTR circles, and 2-LTR circles thought to be the product of autointegration reactions. Recent progress permits the circularization reactions to be studied in vitro. Specific aims include: A. Determination of the optimal conditions for the circularization of viral DNA in vitro. B. Characterization and purification of host-specific proteins required for the formation of 1-LTR and 2-LTR circles. The process of HIV-1 integration is a critical part of the virus life-cycle. Our preliminary studies have opened this area for systematic investigation. Understanding this process may provide opportunities for the development of a novel class of antiviral drugs.

该提议的目的是了解 病毒RNA后的病毒DNA已通过协调转换为DNA 病毒DNA聚合酶和核糖酶活性的作用。 两个 该提案的目标是了解集成的过程 病毒DNA进入宿主DNA并了解病毒的形成过程 DNA圈。 我们的初步工作表明，两者都集成 可以在体外研究循环反应。 目标1。整合反应的表征。 具体目的包括： A.病毒和细胞在 细胞质和核中存在的预一体化复合物 新感染的细胞。 B.使用纯化的有效整合反应重建 病毒和细胞蛋白。 目标2。了解控制病毒形成的反应 DNA圈。 在感染细胞1-LTR圆圈中制造三种类型的病毒DNA圆圈， 简单的2-LTR圆圈和2-LTR圆圈被认为是 自动反应。 最近的进度允许循环 反应在体外研究。 具体目的包括： A.确定最佳条件 体外病毒DNA。 B.所需宿主特异性蛋白的表征和纯化 为1-LTR和2-LTR圆的形成。 HIV-1整合过程是病毒的关键部分 生命周期。 我们的初步研究为系统开放了该领域 调查。 了解此过程可能为 新型抗病毒药物的发展。