BIOCHEMICAL MECHANISMS OF DRUG RESISTANCE IN HIV 1 RT

HIV 1 RT 耐药性的生化机制

• 批准号: 2653878
• 负责人: WALTER A SCOTT
• 金额: $ 26.18万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-15 至 2000-01-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2653878
• 关键词: RNA directed DNA polymerase; chemical kinetics; circular dichroism; deoxyribonucleoside triphosphate; drug resistance; enzyme activity; exonuclease; fluorescence spectrometry; gene mutation; human immunodeficiency virus 1; zidovudine

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.

描述（根据申请人的摘要改编）：调查人员提出 研究AZT电阻的机制和AZT抗性的抑制 通过抑制突变。 他们还很好地研究了改变的RT突变 催化活性，非核苷RT抑制剂和各种核苷酸 类似物。 他们建议定量表征 HIV-1 RT之间形成的构象活性稳定复合物， 底漆 - 培根和脱氧核糖核苷三磷酸（DNTP） 磷酸二酯键合成。 他们很好地监视了 通过 测量核酸酶保护，在没有DNTP的情况下解离速率， 圆形二分性频谱变化和内在蛋白质的变化 荧光。 此外，他们将研究这些突变的影响 在RNase H核酸内切酶和外切核酸酶活性上。 最后，他们很好 表征一种新型的AZTTP依赖性DNA修饰HIV-1 RT的活性 他们最近确定了。 统一的假设是抗药性 突变使酶在DNA链终止时的有效性较低。