INHIBITION OF HUMAN DNA POLYMERASES BY ANTIVIRAL DRUGS

该提案旨在检查抗病毒药物的效果 两种人类DNA聚合酶（α和δ）。涉及 在人类染色体DNA复制中。抗病毒药物对 分离的酶和在其他复制因子存在下 并与它们对重组HIV的影响进行比较 逆转录酶了解这些药物对 人类DNA聚合酶可提供重要的信息， 潜在药物细胞毒性评估和未来设计 抗病毒药物。1. DNA聚合酶a抑制动力学 和DNA聚合酶8的抗病毒药物作用。平行 将进行HIV逆转录酶研究。2.的 DNA聚合酶的抑制机制将在 详细DNA测序凝胶方法和模板引物 寡核苷酸引发的ss-环状DNA的形式将用于分析 所涉及的机制。使用HIV逆转录病毒的平行研究- 将使用模型阿托伐他汀引发的RNA进行转录酶检测 模板结合测序方法学。3.的影响 核苷酸类似物对持续合成能力和链置换的影响 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的一个。4.的 3'至5'核酸外切酶在抑制过程中的特殊作用 将进行研究，特别是在有关链终止 抑制剂;蒙托西肽类似物对该活性的影响将 也要坚决。

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.