Increased Misincorporation Fidelity Observed for Nucleoside Analog Resistance Mutations M184V and E89G in Human Immunodeficiency Virus Type 1 Reverse Transcriptase Does Not Correlate with the Overall Error Rate Measured In Vitro

摘要: Nucleoside analog-resistant variants of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) that displayed higher in vitro polymerase fidelity were previously identified via nucleotide insertion and mispair extension assays. To evaluate the contribution increased primer fidelities on overall error rate HIV-1 RT, we have measured impact two such mutations, E89G M184V, DNA copying an M13 phage-based forward mutation assay. Using this assay, observed frequencies 8.60 × 10−3, 6.26 5.53 12.30 10−3 for wild-type, E89G, double-mutant E89G/M184V RTs, respectively. Therefore, RTs are similar (less than twofold differences) DNA-dependent synthesis. Thus, rather large increases deoxynucleoside triphosphate appear not to influence these mutants. However, a qualitative analysis mutations induced revealed significant differences mutational spectra between wild-type mutant enzymes.