Efficiency of extension of mismatched primer termini across from cisplatin and oxaliplatin adducts by human DNA polymerases beta and eta in vitro.
DNA polymerases beta and eta are among the few eukaryotic polymerases known to efficiently bypass cisplatin and oxaliplatin adducts in vitro. Our laboratory has previously established that both polymerases misincorporated dTTP with high frequency across from cisplatin- and oxaliplatin-GG adducts. This decrease in polymerase fidelity on platinum-damaged DNA could lead to in vivo mutations, if this base substitution were efficiently elongated. In this study, we performed a steady-state kinetic analysis of the steps required for fixation of dTTP misinsertion during translesion synthesis past cisplatin- and oxaliplatin-GG adducts by pol beta and pol eta. The efficiency of translesion synthesis by pol eta past Pt-GG adducts was very similar to that observed for this polymerase when the template contains thymine-thymine dimers. This finding suggested that pol eta could play a role in translesion synthesis past platinum-GG adducts in vivo. On the other hand, translesion synthesis past platinum-GG adducts by pol beta was much less efficient. Translesion synthesis by pol eta is likely to be predominantly error-free, since the probability of correct insertion and extension by pol eta was 1000-2000-fold greater than the probability of incorrect insertion and extension. Our results also indicated that for pol eta the frequency of misincorporation is the same across from the 3'G and the 5'G of the platinum-GG adducts for both cisplatin and oxaliplatin adducts. On the other hand, pol beta is more likely to misinsert at the 3'G of the adducts and misinsertion occurs at higher frequency for oxaliplatin-GG than for cisplatin-GG adducts.
Bassett
Ekaterina
E
Department of Biochemistry and Biophysics, Lineberger Comprehensive Cancer Center, School of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599-7260, USA.
Vaisman
Alexandra
A
Havener
Jody M
JM
Masutani
Chikahide
C
Hanaoka
Fumio
F
Chaney
Stephen G
SG
D020137:Base Pair Mismatch; D002945:Cisplatin / Q000737:chemistry*; D018736:DNA Adducts / Q000737:chemistry*; D004249:DNA Damage; D019951:DNA Polymerase beta / Q000737:chemistry*; D017931:DNA Primers / Q000737:chemistry*; D004260:DNA Repair; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry*; D006801:Humans; D007700:Kinetics; D009944:Organoplatinum Compounds / Q000737:chemistry*; D000077150:Oxaliplatin; D011994:Recombinant Proteins / Q000737:chemistry; D013698:Templates, Genetic; D013942:Thymine Nucleotides / Q000737:chemistry; D000099270:Y-Family DNA Polymerases
D002945:Cisplatin; D018736:DNA Adducts; D019951:DNA Polymerase beta; D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase; D009944:Organoplatinum Compounds; D000077150:Oxaliplatin; D011994:Recombinant Proteins; D013942:Thymine Nucleotides; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases; C024157:thymidine 5'-triphosphate