BIOCHEMICAL MECHANISMS OF DRUG RESISTANCE IN HIV 1 RT

描述（改编自申请人的摘要）：研究人员建议 研究AZT耐药机制和抑制AZT耐药 通过抑制突变。 他们还研究了改变 RT 突变 催化活性、非核苷类RT抑制剂和各种核苷酸 类似物。 他们建议定量地表征 HIV-1 RT 之间形成的构象活性稳定复合物， 引物模板和脱氧核糖核苷三磷酸 (dNTP) 磷酸二酯键的合成被阻止。 他们很好地监控 稳定复合物形成过程中发生的结构变化 测量核酸酶保护、没有 dNTP 时的解离率， 圆二色性光谱变化和内在蛋白质的变化 荧光。 此外，他们还将研究这些突变的影响 RNase H 核酸内切酶和核酸外切酶活性。 最后他们还好 描述了 HIV-1 RT 的新型 AZTTP 依赖性 DNA 修饰活性， 他们最近发现。 统一的假设是耐药性 突变使酶在 DNA 链终止时的效率降低。

DESCRIPTION (Adapted from Applicant's Abstract): The investigators propose to study the mechanism of AZT resistance and suppression of AZT resistance by suppressor mutations. They well also study RT mutations that alter catalytic activity, non-nucleoside RT inhibitors, and various nucleotide analogues. They propose to characterize quantitatively the formation of conformationally active stable complex formed between HIV-1 RT, primer-template, and deoxyribonucleoside triphosphate (dNTP) when phosphodiester bond synthesis is prevented. They well monitor the formational changes that occur in the formation of stable complex by measuring nuclease protection, dissociation rate in absence of dNTP, circular dichroism spectral changes, and changes in intrinsic protein fluorescence. In addition, they will study the effects of these mutations on RNase H endonuclease and exonuclease activity. Finally, they well characterize a novel AZTTP-dependent DNA modifying activity of HIV-1 RT that they recently identified. The unifying hypothesis is that drug-resistance mutations render the enzyme less effective at DNA chain termination.