Mutations in HIV-1 Reverse Transcriptase Affect the Errors Made in a Single Cycle of Viral Replication
Mutations in HIV-1 Reverse Transcriptase Affect the Errors Made in a Single Cycle of Viral Replication
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DOI：
10.1128/jvi.00302-14
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发表时间：
2014-07-01
影响因子：
5.4
通讯作者：
Hughes, Stephen H.
中科院分区：
文献类型：
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作者：
Abram, Michael E.;Ferris, Andrea L.;Hughes, Stephen H.
患者体内HIV-1的遗传变异是由于病毒复制率高、病毒载量高以及病毒复制过程中出现的错误。逆转录酶（RT）中改变脱氧核苷三磷酸（dNTP）结合口袋的一些突变，包括那些赋予核苷/核苷酸类似物抗性的突变，会影响复制过程中的dNTP选择。RT突变对体内错误谱（性质、位置和频率）的影响知之甚少。我们先前确定了不同类型突变的突变率和频率，并确定了lacZ α突变的热点。（lacZ的α互补区）报告基因由HIV-1载体携带，使用野生型RT进行复制。（Y115F，M184V，M184I，和Q151 M），对HIV-1总体突变率的影响相对较小（与野生型相比小于3倍）显著增加了lacZ α报告基因中某些特定位置的突变。我们还表明，侧翼的报告基因的序列的变化可以影响报告中出现的突变。这些数据表明，HIV-1 RT或核酸模板序列的变化可以影响病毒复制过程中产生的突变谱。这可能会影响患者体内耐药突变体和免疫逃逸突变体的产生。
The genetic variation in HIV-1 in patients is due to the high rate of viral replication, the high viral load, and the errors made during viral replication. Some of the mutations in reverse transcriptase (RT) that alter the deoxynucleoside triphosphate (dNTP)binding pocket, including those that confer resistance to nucleoside/nucleotide analogs, affect dNTP selection during replication. The effects of mutations in RT on the spectrum (nature, position, and frequency) of errors made in vivo are poorly understood. We previously determined the mutation rate and the frequency of different types of mutations and identified hot spots for mutations in a lacZ alpha (the alpha complementing region of lacZ) reporter gene carried by an HIV-1 vector that replicates using wild-type RT. We show here that four mutations (Y115F, M184V, M184I, and Q151M) in the dNTP-binding pocket of RT that had relatively small effects on the overall HIV-1 mutation rate (less than 3-fold compared to the wild type) significantly increased mutations at some specific positions in the lacZ alpha reporter gene. We also show that changes in a sequence that flanks the reporter gene can affect the mutations that arise in the reporter. These data show that changes either in HIV-1 RT or in the sequence of the nucleic acid template can affect the spectrum of mutations made during viral replication. This could, by implication, affect the generation of drug-resistant mutants and immunological-escape mutants in patients.