DNA polymerase beta bypasses in vitro a single d(GpG)-cisplatin adduct placed on codon 13 of the HRAS gene.
We have examined the capacity of calf thymus DNA polymerases alpha, beta, delta, and epsilon to perform in vitro translesion synthesis on a substrate containing a single d(GpG)-cisplatin adduct placed on codon 13 of the human HRAS gene. We found that DNA synthesis catalyzed by DNA polymerases alpha, delta, and epsilon was blocked at the base preceding the lesion. Addition of proliferating cell nuclear antigen to DNA polymerase delta and replication protein A to DNA polymerase alpha did not restore their capacity to elongate past the adduct. On the other hand, DNA polymerase beta efficiently bypassed the cisplatin adduct. Furthermore, we observed that DNA polymerase beta was the only polymerase capable of primer extension of a 3'-OH located opposite the base preceding the lesion. Likewise, DNA polymerase beta was able to elongate the arrested replication products of the other three DNA polymerases, thus showing its capacity to successfully compete with polymerases alpha, delta, and epsilon in the stalled replication complex. Our data suggest (i) a possible mechanism enabling DNA polymerase beta to bypass a d(GpG)-cisplatin adduct in vitro and (ii) a role for this enzyme in processing DNA damage in vivo.
Hoffmann
J S
JS
Laboratoire de Pharmacologie et Toxicologie Fondamentales, Centre National de la Recherche Scientifique, Toulouse, France.
Pillaire
M J
MJ
Maga
G
G
Podust
V
V
Hübscher
U
U
Villani
G
G
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002945:Cisplatin / Q000494:pharmacology*; D003062:Codon; D004256:DNA Polymerase I / Q000378:metabolism*; D015226:Dinucleoside Phosphates / Q000494:pharmacology*; D011905:Genes, ras; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D013950:Thymus Gland / Q000201:enzymology
D003062:Codon; D015226:Dinucleoside Phosphates; D009838:Oligodeoxyribonucleotides; C094059:cisplatin-deoxy(guanosine monophosphate guanosine) adduct; D004256:DNA Polymerase I; D002945:Cisplatin