Fidelity of HIV-1 reverse transcriptase.
The human immunodeficiency virus type 1 (HIV-1) shows extensive genetic variation and undergoes rapid evolution. The fidelity of purified HIV-1 reverse transcriptase was measured during DNA polymerization in vitro by means of three different assays. Reverse transcriptase from HIV-1 introduced base-substitution errors in DNA from the bacteriophage phi X174 amber3 at estimated frequencies of 1/2000 to 1/4000. Analyses of misincorporation rates opposite a single template adenine residue showed that HIV-1 reverse transcriptase catalyzed nucleotide mismatches with a specificity of A:C much greater than A:G greater than A:A. The high error rate of HIV-1 reverse transcriptase in vitro translates to approximately five to ten errors per HIV-1 genome per round of replication in vivo. This high error rate suggests that misincorporation by HIV-1 reverse transcriptase is, at least in part, responsible for the hypermutability of the AIDS virus. The specificity of misincorporation may provide a basis for the systematic construction of antiviral nucleosides.
Preston
B D
BD
Department of Pathology, University of Washington, Seattle 98195.
Poiesz
B J
BJ
Loeb
L A
LA
D009189:Avian Myeloblastosis Virus / Q000201:enzymology; D010584:Bacteriophage phi X 174 / Q000235:genetics; D004247:DNA / Q000096:biosynthesis*; D004257:DNA Polymerase II / Q000378:metabolism; D004279:DNA, Viral / Q000096:biosynthesis; D004591:Electrophoresis, Polyacrylamide Gel; D006678:HIV / Q000201:enzymology* / Q000235:genetics; D007700:Kinetics; D008979:Moloney murine leukemia virus / Q000201:enzymology; D009154:Mutation; D009711:Nucleotides / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*
D004279:DNA, Viral; D009711:Nucleotides; D004247:DNA; D012194:RNA-Directed DNA Polymerase; D004257:DNA Polymerase II