The efficiency and fidelity of translesion synthesis past cisplatin and oxaliplatin GpG adducts by human DNA polymerase beta.
DNA polymerase beta (pol beta) is the only mammalian DNA polymerase identified to date that can catalyze extensive bypass of platinum-DNA adducts in vitro. Previous studies suggest that DNA synthesis by pol beta is distributive on primed single-stranded DNA and processive on gapped DNA. The data presented in this paper provide an analysis of translesion synthesis past cisplatin- and oxaliplatin-DNA adducts by pol beta functioning in both distributive and processive modes using primer extension and steady-state kinetic experiments. Translesion synthesis past Pt-DNA adducts was greater with gapped DNA templates than with single-stranded DNA templates. In the processive mode pol beta did not discriminate between cisplatin and oxaliplatin adducts, while in the distributive mode it displayed about 2-fold increased ability for translesion synthesis past oxaliplatin compared with cisplatin adducts. The differentiation between cisplatin and oxaliplatin adducts resulted from a K(m)-mediated increase in the efficiency of dCTP incorporation across from the 3'-G of oxaliplatin-GG adducts. Rates of misincorporation across platinated guanines determined by the steady-state kinetic assay were higher in reactions with primed single-stranded templates than with gapped DNA and a slight increase in the misincorporation of dTTP across from the 3'-G was found for oxaliplatin compared with cisplatin adducts.
Vaisman
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A
Department of Biochemistry and Biophysics, Lineberger Comprehensive Cancer Center, School of Medicine, University of North Carolina, Chapel Hill 27599-7260, USA.
Chaney
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SG
D000970:Antineoplastic Agents / Q000494:pharmacology*; D001483:Base Sequence; D002945:Cisplatin / Q000494:pharmacology*; D018736:DNA Adducts / Q000378:metabolism*; D019951:DNA Polymerase beta / Q000378:metabolism*; D017931:DNA Primers / Q000378:metabolism; D004260:DNA Repair; D004261:DNA Replication; D003845:Deoxycytosine Nucleotides / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009944:Organoplatinum Compounds / Q000494:pharmacology*; D000077150:Oxaliplatin; D011994:Recombinant Proteins / Q000378:metabolism; D013942:Thymine Nucleotides / Q000378:metabolism; D013997:Time Factors
D000970:Antineoplastic Agents; D018736:DNA Adducts; D017931:DNA Primers; D003845:Deoxycytosine Nucleotides; D009944:Organoplatinum Compounds; D011994:Recombinant Proteins; D013942:Thymine Nucleotides; D000077150:Oxaliplatin; C024107:2'-deoxycytidine 5'-triphosphate; D019951:DNA Polymerase beta; D002945:Cisplatin; C024157:thymidine 5'-triphosphate