Fidelity of in vitro DNA strand transfer reactions catalyzed by HIV-1 reverse transcriptase.
The fidelity of DNA strand transfer reactions catalyzed by human immunodeficiency virus type 1 reverse transcriptase has been studied in vitro. A model system involving two sequential DNA strand transfers was developed to simulate the process of forced copy-choice recombination. A propensity for nucleotide misincorporation at the junction of the strand transfer, as determined by DNA sequencing of the reaction products, was found consistent with a model involving the addition of nontemplate-directed nucleotides prior to the transfer of nascent DNA onto the accepting RNA template [Peliska, J. A., & Benkovic, S. J. (1992) Science 258, 1112]. The kinetic and mechanistic factors that may dictate which nucleotide bases are incorporated at recombination sites during strand transfer and the possible consequences of recombination-induced mutagenesis in vivo are discussed.
Peliska
J A
JA
Department of Chemistry, Pennsylvania State University, Wartik Laboratory, University Park 16802.
Benkovic
S J
SJ
D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D016296:Mutagenesis; D012313:RNA / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D011995:Recombination, Genetic; D013698:Templates, Genetic
D012313:RNA; D004247:DNA; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase