Unselected mutations in the human immunodeficiency virus type 1 genome are mostly nonsynonymous and often deleterious
Unselected mutations in the human immunodeficiency virus type 1 genome are mostly nonsynonymous and often deleterious
复制标题
DOI：
10.1128/jvi.78.5.2426-2433.2004
复制
发表时间：
2004-03-01
影响因子：
5.4
通讯作者：
Hui, HX
中科院分区：
文献类型：
--
作者：
Gao, F;Chen, Y;Hui, HX
使用纯化的逆转录酶或单轮感染系统估算了 1 型人类免疫缺陷病毒 (HIV-1) 基因组的突变率。由于使用小序列作为模板，因此只能推断总体突变率，并且突变的生物学意义未知。为了直接估计HIV-1突变率并了解突变的潜在生物学影响，我们利用λ噬菌体文库方法和长程PCR技术从单轮感染的淋巴细胞贴壁细胞中获得了19个完整或接近全长的原病毒基因组。对160,000条序列的分析表明，HIV-1基因组的总体突变率为每个复制周期每个碱基5.4 X 10(-5)。在一个感染周期内，每个病毒基因组平均产生 1.1 个突变（范围为 0 到 3 个）。对 HIV-1 基因组突变的检查表明，蛋白质编码区内的所有位点突变都是非同义突变。在所有突变中，一半是有害的（过早终止密码子和缺失）并会导致基因组缺陷。通过将相同的系统应用于逆转录酶拇指区域具有 G262A 突变的 HIV-1 基因组，观察到删除和插入突变率显着增加，但没有发现病毒基因组的总体突变率增加。
Mutation rates of human immunodeficiency virus type 1 (HIV-1) genomes have been estimated using purified reverse transcriptase or single-round infection system. Since small sequences were used as templates, the overall mutation rates could only be extrapolated and the biological significance of mutations is unknown. For direct estimation of HIV-1 mutation rates and understanding of the potential biological influences of mutations, we obtained 19 complete or nearly full-length proviral genomes from single-round-infected adherent cells of lymphocytes by using a lambda phage library method and a long-range PCR technique. Analysis of 160,000 lip of sequences showed that the overall mutation rate of HIV-1 genomes was 5.4 X 10(-5) per base per replication cycle. On average, 1.1 mutations (range, 0 to 3) were generated in each viral genome during one infection cycle. Inspection of the mutations in the HIV-1 genome revealed that all site mutations within protein-coding regions were nonsynonymous mutations. Among all mutations, half were deleterious (premature stop codon and deletions) and would result in defective genomes. By applying the same system to an HIV-1 genome with a G262A mutation in the thumb region of the reverse transcriptase, a significant increase was observed in deletion and insertion mutation rates but no increase in the overall mutation rate in viral genomes was found.