BIOCHEMICAL MECHANISMS OF DRUG RESISTANCE IN HIV 1 RT

HIV 1 RT 耐药性的生化机制

• 批准号: 2004810
• 负责人: WALTER A SCOTT
• 金额: $ 25.6万
• 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/2004810
• 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（改编自申请人摘要）：研究人员建议 研究AZT耐药的机制及抑制AZT耐药的方法 抑制基因突变。 他们还研究了RT突变， 催化活性、非核苷RT抑制剂和各种核苷酸 类似物 他们建议定量描述 HIV-1 RT之间形成构象活性稳定的复合物， 引物模板和脱氧核苷三磷酸（dNTP）时 磷酸二酯键的合成被阻止。 他们很好地监测了 在形成稳定复合物时发生的结构变化， 测量核酸酶保护，在不存在dNTP的情况下的解离速率， 圆二色光谱的变化，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 对RNase H核酸内切酶和核酸外切酶活性的影响。 最后，他们 表征了HIV-1 RT的新的AZTTP依赖性DNA修饰活性， 他们最近发现。 统一的假设是抗药性 突变使酶在DNA链终止时不那么有效。