PHARMACOLOGY OF THE HIV VIRAL DNA

HIV 病毒 DNA 的药理学

• 批准号: 3838171
• 负责人: Y POMMIER
• 金额: --
• 依托单位: DIVISION OF CANCER TREATMENT
• 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/3838171
• 关键词: DNA topoisomerases; chloroquine; enzyme mechanism; genetic mapping; genetic promoter element; genetic recombination; genetic regulatory element; human immunodeficiency virus; nucleic acid repetitive sequence; nucleic acid sequence; oligonucleotides; transcription factor; virus DNA; virus genetics; virus infection mechanism; virus replication

Each step of the Human Immunodeficiency Virus (HIV) replication cycle represents a potential target for therapeutic intervention. A critical step is integration of the viral DNA, synthesized by reverse transcriptase within the cytoplasm of an infected cell, into the host genome. We have set up an in vitro retroviral DNA integration system using recombinant HIV integrase and an oligonucleotide which corresponds to the U5 end of HIV DNA. Two steps of the integration reaction can be analyzed in the assay: (i) nucleolytic cleavage which removes 2 nucleotides from the 3'ends of the double-stranded DNA, (ii) DNA strand transfer which couples the joining of the viral DNA into the target DNA with cleavage of the target DNA at the site of insertion. Using this assay, we have identified several groups of drugs that effectively inhibit the integration reaction including some DNA intercalators such as chloroquine which are active at micromolar concentrations. Interestingly, inhibitors of DNA topoisomerases I and II are generally inactive. Our current effort is aimed at investigating several classes of non-DNA binders which could be tested for integrase inhibition in vivo. Long Terminal Repeats (LTRs) are repeated DNA sequences at both ends of retroviral DNA. Besides their role as HIV integrase targets, the LTRs are the promoter regions of HIV DNA. They contain well defined transcription regulatory elements which are critical for making new viral RNA and proteins. Because of our recent finding that DNA topoisomerase II has preferential sites within the promoter region of the human MYC proto-oncogene (Pommier et al., Cancer Res. 1992;52:3125-30) and because topoisomerase I activity appears to facilitate transcription, we are mapping the interactions of DNA topoisomerases with the LTR regions and studying the effects of various topoisomerase inhibitors on the LTRs.

人类免疫缺陷病毒（HIV）复制周期的每一步 代表治疗干预的潜在靶点。 一个关键 步骤是整合病毒DNA，通过逆转录合成， 受感染细胞的细胞质内的转录酶，进入宿主 基因组 我们建立了体外逆转录病毒DNA整合系统 使用重组HIV整合酶和相应的寡核苷酸， HIV DNA的U 5末端 积分反应的两个步骤可以是： 在测定中分析：（i）溶核裂解，其去除2 来自双链DNA的3 '末端的核苷酸，（ii）DNA链 将病毒DNA的连接偶联到靶DNA中的转移 在插入位点切割靶DNA。 使用此 通过分析，我们已经确定了几组药物， 抑制整合反应，包括一些DNA嵌入剂， 氯喹在微摩尔浓度下具有活性。 有趣的是，DNA拓扑异构酶I和II的抑制剂通常是 不活跃。 我们目前的努力旨在调查几个类 的非DNA结合剂，其可用于测试整合酶抑制， vivo. 长末端重复序列（LTR）是在DNA的两端重复的DNA序列。 逆转录病毒DNA。 除了作为HIV整合酶靶点的作用外， 是HIV DNA的启动子区域。 它们包含定义明确的 转录调控元件对于制造新的病毒至关重要， RNA和蛋白质。 因为我们最近发现DNA拓扑异构酶 II在人MYC的启动子区域内具有优先位点 原癌基因（Pomelians等，Cancer Res. 1992;52：3125-30），并且因为 拓扑异构酶I活性似乎促进转录，我们是 绘制DNA拓扑异构酶与LTR区域的相互作用， 研究各种拓扑异构酶抑制剂对LTR的影响。