In vitro bypass replication of the cisplatin-d(GpG) lesion by calf thymus DNA polymerase beta and human immunodeficiency virus type I reverse transcriptase is highly mutagenic.
Eukaryotic DNA polymerase beta and the reverse transcriptases are the most inaccurate of the known DNA polymerases. We report here mutagenic replication in vitro past intrastrand N(7)G-N(7)G chelates of the cis-diamminedichloroplatinum(II), the major DNA adduct of the antitumor agent cisplatin by calf thymus DNA polymerase beta and human immunodeficiency virus type I reverse transcriptase (42% and 26% mutations, respectively). The most frequent modifications generated by both enzymes were one-base frameshift deletions. Only one mutational hot spot opposite the platinated guanines was observed with human immunodeficiency virus type I reverse transcriptase, while two hot spots were generated by DNA polymerase beta, one at the base situated 5' to the lesion and the other situated 4-6 nucleotides 5' to the adduct. An unusual mutagenic event, tandem replication of a 12-base pair sequence, was observed with DNA polymerase beta. The mutational spectra of the two DNA polymerases suggest that template slippage occurred with higher frequency in the presence of the more distributive DNA polymerase beta.
Hoffmann
J S
JS
Institut de Pharmacologie et de Biologie Structurale, CNRS, 205, route de Narbonne, 31077 Toulouse cedex, France.
Pillaire
M J
MJ
Garcia-Estefania
D
D
Lapalu
S
S
Villani
G
G
D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry; D001483:Base Sequence; D002417:Cattle; D002474:Cell-Free System; D002945:Cisplatin / Q000737:chemistry*; D018736:DNA Adducts; D004256:DNA Polymerase I / Q000378:metabolism*; D004261:DNA Replication; D054303:HIV Reverse Transcriptase; D008969:Molecular Sequence Data; D016296:Mutagenesis; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D017384:Sequence Deletion
D000970:Antineoplastic Agents; D018736:DNA Adducts; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; D004256:DNA Polymerase I; D002945:Cisplatin