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- W3122502631 abstract "DNA polymerases insert dATP opposite the oxidative damage product 7,8 dihydro-8-oxodeoxyguanosine (8-oxoG) instead of dCTP, to the extent of > 90% with some polymerases. Steady-state kinetics with the Y-family Sulfolobus solfataricus DNA polymerase IV (Dpo4) showed 25-fold higher incorporation efficiency of dCTP > dATP opposite 8-oxoG and 4-fold higher efficiency of extension beyond an 8-oxoG:C pair than an 8-oxoG:A pair. The catalytic efficiency for these events with dCTP was similar for G and 8-oxoG templates. Mass spectral analysis of extended DNA primers showed ≥ 95% incorporation of dCTP > dATP opposite 8-oxoG. Pre-steady-state kinetics showed faster rates of dCTP incorporation opposite 8-oxoG than G. The measured Kd,dCTP was 15-fold lower for an oligonucleotide containing 8-oxoG than with G. Extension beyond an 8-oxoG:C pair was similar to G:C and faster than for an 8-oxoG:A pair, in contrast to other polymerases. The Ea for dCTP insertion opposite 8-oxoG was lower than opposite G. Crystal structures of Dpo4 complexes with oligonucleotides were solved with C, A, and G nucleoside triphosphates placed opposite 8-oxG. With ddCTP, dCTP, and dATP the phosphodiester bonds were formed even in the presence of Ca2+. The 8-oxoG:C pair showed classic Watson-Crick geometry; the 8-oxoG:A pair was in the syn:anti configuration, with the A hybridized in a Hoogsteen pair with the 8-oxoG. With dGTP placed opposite 8-oxoG, pairing was not to the 8-oxoG but to the 5′ C (and in classic Watson-Crick geometry), consistent with the low frequency of this frameshift event observed in the catalytic assays." @default.
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- W3122502631 date "2006-03-01" @default.
- W3122502631 modified "2023-10-18" @default.
- W3122502631 title "Efficient and High Fidelity Incorporation of dCTP Opposite 7,8‐Dihydro‐8‐oxodeoxyguanosine by Sulfolobus solfataricus DNA Polymerase Dpo4" @default.
- W3122502631 doi "https://doi.org/10.1096/fasebj.20.4.a39-c" @default.
- W3122502631 hasPublicationYear "2006" @default.
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