Catalytic efficiency and phenotype of HIV-1 proteases encoding single critical resistance substitutions
Catalytic efficiency and phenotype of HIV-1 proteases encoding single critical resistance substitutions
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DOI：
10.1006/viro.2002.1520
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发表时间：
2002-08-15
期刊：
影响因子：
3.7
通讯作者：
Martínez, MA
中科院分区：
文献类型：
--
作者：
Cabana, M;Fernàndez, G;Martínez, MA
我们已经表明，噬菌体λ基因筛选系统可能是有用的预测HIV-1蛋白酶的活性和表型的过程中的病毒感染和抗逆转录病毒治疗。这种简单而快速的遗传筛选系统已被用于表征编码单一初级抗性取代的HIV-1蛋白酶。除了在位置46和84处具有氨基酸变化的蛋白酶之外，含有单抗性取代的蛋白酶显示出比WT酶更低的催化效率。单个突变体可以通过它们的效率来鉴定，表明蛋白酶活性的适度差异可以用这种简单的测定来监测。总体而言，药物敏感性可以通过引入单一突变来降低。然而，高水平的蛋白酶抑制剂（PI）的耐药性，只有实现了多个突变的蛋白酶。单个突变体显示的耐药性小幅但可重复的增加也证明了这种遗传筛查系统检测药物敏感性小幅降低的能力。这些结果表明，这里使用的噬菌体λ基因筛选系统是一个有用的工具，在分析的特定贡献的突变在HIV蛋白酶编码区或在特定的切割位点，影响PI抗性的过程。(C)2002 Elsevier Science（美国）。
We have shown that a bacteriophage lambda genetic screen system may be useful in predicting the activity and phenotype of HIV-1 protease in the course of viral infection and antiretroviral therapy. This simple and rapid genetic screening system has been used here to characterize HIV-1 proteases encoding single primary resistance substitutions. Except for proteases with amino acid changes at positions 46 and 84, proteases containing single-resistance substitutions displayed a lower catalytic efficiency than the WT enzyme. Single mutants could be identified by their efficiency, demonstrating that modest differences in protease activity can be monitored with this simple assay. Overall, drug susceptibility could be reduced by introduction of single mutations. However, high-level protease inhibitor (PI) resistance was only achieved by multiple mutated proteases. The small but reproducible increase in resistance displayed by single mutants also demonstrated the ability of this genetic screen system for detecting minor reductions in drug susceptibility. These results show that the bacteriophage lambda genetic screen system used here is a useful tool in the analysis of specific contribution of mutations in the HIV protease-coding region or in specific cleavage sites that affect the process of PI resistance. (C) 2002 Elsevier Science (USA).