Involvement of specialized DNA polymerases in mutagenesis by 8-hydroxy-dGTP in human cells.
The mutagenicity of an oxidized form of dGTP, 8-hydroxy-2'-deoxyguanosine 5'-triphosphate (8-OH-dGTP), was examined using human 293T cells. Shuttle plasmid DNA containing the supF gene was first transfected into the cells, and then 8-OH-dGTP was introduced by means of osmotic pressure. The DNAs replicated in the cells were recovered and then transfected into Escherichia coli. 8-OH-dGTP induced A:T-->C:G substitution mutations in the cells. The knock-downs of DNA polymerases eta and zeta, and REV1 by siRNAs reduced the A:T-->C:G substitution mutations, suggesting that these DNA polymerases are involved in the misincorporation of 8-OH-dGTP opposite A in human cells. In contrast, the knock-down of DNA polymerase iota did not affect the 8-OH-dGTP-induced mutations. The decrease in the induced mutation frequency was more evident by double knock-downs of DNA pols eta plus zeta and REV1 plus DNA pol zeta (but not by that of DNA pol eta plus REV1), suggesting that REV1-DNA pol eta and DNA pol zeta work in different steps. These results indicate that specialized DNA polymerases are involved in the mutagenesis induced by the oxidized dGTP.
Satou
Kazuya
K
Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
Hori
Mika
M
Kawai
Kazuaki
K
Kasai
Hiroshi
H
Harashima
Hideyoshi
H
Kamiya
Hiroyuki
H
D000080242:8-Hydroxy-2'-Deoxyguanosine; D002460:Cell Line; D002478:Cells, Cultured; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D003848:Deoxyguanine Nucleotides / Q000494:pharmacology*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000378:metabolism; D016296:Mutagenesis; D009154:Mutation / Q000235:genetics*; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D019384:Nucleic Acid Synthesis Inhibitors; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D000094769:DNA Polymerase iota; D000099270:Y-Family DNA Polymerases
D000080242:8-Hydroxy-2'-Deoxyguanosine; D004259:DNA-Directed DNA Polymerase; D003848:Deoxyguanine Nucleotides; D009687:Nuclear Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D009713:Nucleotidyltransferases; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000094769:DNA Polymerase iota; C521293:8-hydroxy-2'-deoxyguanosine 5'-triphosphate; C425929:DNA polymerase zeta; C429004:REV1 protein, human; D000099270:Y-Family DNA Polymerases