BIOCHEMICAL MECHANISMS OF DRUG RESISTANCE IN HIV 1 RT

描述(改编自申请者摘要)：调查人员建议 AZT耐药机制及抑制AZT耐药的研究 通过抑制子突变。他们还很好地研究了RT突变 催化活性、非核苷类逆转录酶抑制剂和各种核苷酸 类似物。他们建议定量地描述 在HIV-1RT， 引物-模板和脱氧核糖核苷三磷酸(DNTP) 阻止了磷酸二酯键的合成。他们很好地监控着 在稳定络合物的形成过程中发生的构型变化 测定核酸酶保护，无dNTP时的离解率， 圆二色谱的变化，以及内在蛋白质的变化 荧光。此外，他们还将研究这些突变的影响。 核糖核酸酶H内切酶和核酸外切酶活性的研究最后，他们很好地 HIV-1逆转录酶依赖于AZTTP的DNA修饰活性 他们最近发现。统一的假设是耐药 突变使该酶在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.