INHIBITION OF HUMAN DNA POLYMERASES BY ANTIVIRAL DRUGS

该提案旨在检查抗病毒药物的效果。 在人类的两个DNA聚合酶(α和Delta)上。都牵涉其中 在人类染色体DNA复制中。抗病毒药物对人类免疫功能的影响 分离的酶以及在其他复制因子存在的情况下 将被检测并比较它们对重组HIV的影响 逆转录酶。了解这类药物对健康的影响 人类DNA聚合酶可能提供与以下相关的重要信息 潜在药物细胞毒性的评估和未来的设计 抗病毒药物。1.抑制DNA聚合酶a的动力学 以及由抗病毒药物引起的DNA聚合酶8的研究。平行 将进行艾滋病毒逆转录酶的研究。2. DNA聚合酶的抑制机制将在 细节。DNA测序凝胶方法和模型模板-引物法 寡核苷酸引发的单链环状DNA将用于分析 所涉及的机制。使用HIV反向的平行研究- 转录酶将使用模型寡核苷酸启动的RNA来执行 模板与测序方法相结合。3. 核苷酸类似物对加工性和链移位的影响 将通过使用模型定义的底物来研究反应， 包括携带拓扑稳定的复制叉子的一个。4. 3‘到5’核酸外切酶在抑制过程中的特殊作用 将被研究，特别是与链终止有关的 抑制剂；单核苷酸类似物对这种活性的影响将 也要有决心。

The proposal is directed toward examining the effects of antiviral drugs on the two human DNA polymerases (alpha and delta) which. are involved in human chromosomal DNA replication. The effects of antiviral drugs on the isolated enzymes and in the presence of other replication factors will be examined and compared to their effects on recombinant HIV reverse transcriptase. A knowledge of the effects of such drugs on the human DNA polymerases may provide important inforrnation in relation to the assessment of potential drug cytotoxicity and for the future design of antiviral drugs. 1. The kinetics of inhibition of DNA polymerase a and DNA polymerase 8 by antiviral drugs will be studied. Parallel studies with HIV reverse transcriptase will be performed. 2. The mechanisms of the inhibition of the DNA polymerases will be studied in detail. DNA sequencing gel methods and model template-primers in the form of oligonucleotide primed ss-circular DNA will be used to analyze the mechanisms involved. Parallel studies using HIV reverse- transcriptase will be performed using model oligonucleotide-primed RNA templates in combination with sequencing methodology. 3. The effects of nucleotide analogs on processivity and on the strand displacement reactions will be studied by the use of model defined substrates, including one carrying a topologically stable replication fork. 4. The particular role of the 3' to 5' exonuclease in the inhibition processes will be studied, particularly in relation to chain-terminating inhibitors; the effects of mononucleotide analogs on this activity will also be determined.