Increased polymerase fidelity of the 3TC-resistant variants of HIV-1 reverse transcriptase

被引:73
作者
Essink, BBO [1 ]
Back, NKT [1 ]
Berkhout, B [1 ]
机构
[1] UNIV AMSTERDAM, ACAD MED CTR, DEPT HUMAN RETROVIROL, NL-1100 DE AMSTERDAM, NETHERLANDS
关键词
D O I
10.1093/nar/25.16.3212
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Human immunodeficiency virus type 1 (HIV-1) variants with resistance mutations in the reverse transcriptase (RT) gene appear during drug therapy with the nucleoside analogue 2',3'-dideoxy-3'-thiacytidine (3TC). These STC-resistant RT variants have a single point mutation that changes the 184Met residue into either Val or Ile. Both codon 184 variants are frequently observed in 3TC-treated patients and can also be selected in cell culture infections. We demonstrated previously that the 184Ile and 184Val RT enzymes exhibit a processivity defect in in vitro assays, with 184Ile being the least processive enzyme [Met(wt) > Val. lie]. In this study, we measured the polymerase fidelity of the wild-type (184Met) and 3TC-resistant RT enzymes (184Ile and 184Val) on DNA and RNA templates. Both virion-extracted and Escherichia coli-expressed recombinant RT enzymes were used to measure the nucleotide misinsertion and mispair extension efficiencies. The 3TC-resistant enzymes were more accurate than the wild-type RT protein in both type of assays. The order of accuracy observed for the codon 184 variants [lie > Val > Met(wt)] may suggest an inverse correlation between the fidelity and processivity properties of these enzymes.
引用
收藏
页码:3212 / 3217
页数:6
相关论文
共 42 条