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The abundant DNA adduct N 7-methyl deoxyguanosine contributes to miscoding during replication by human DNA polymerase ?.


ABSTRACT: Aside from abasic sites and ribonucleotides, the DNA adduct N 7-methyl deoxyguanosine (N7 -CH3 dG) is one of the most abundant lesions in mammalian DNA. Because N7 -CH3 dG is unstable, leading to deglycosylation and ring-opening, its miscoding potential is not well-understood. Here, we employed a 2'-fluoro isostere approach to synthesize an oligonucleotide containing an analog of this lesion (N7 -CH3 2'-F dG) and examined its miscoding potential with four Y-family translesion synthesis DNA polymerases (pols): human pol (hpol) ?, hpol ?, and hpol ? and Dpo4 from the archaeal thermophile Sulfolobus solfataricus We found that hpol ? and Dpo4 can bypass the N7 -CH3 2'-F dG adduct, albeit with some stalling, but hpol ? is strongly blocked at this lesion site, whereas hpol ? showed no distinction with the lesion and the control templates. hpol ? yielded the highest level of misincorporation opposite the adduct by inserting dATP or dTTP. Moreover, hpol ? did not extend well past an N 7-CH3 2'-F dG:dT mispair. MS-based sequence analysis confirmed that hpol ? catalyzes mainly error-free incorporation of dC, with misincorporation of dA and dG in 5-10% of products. We conclude that N 7-CH3 2'-F dG and, by inference, N 7-CH3 dG have miscoding and mutagenic potential. The level of misincorporation arising from this abundant adduct can be considered as potentially mutagenic as a highly miscoding but rare lesion.

SUBMITTER: Njuma OJ 

PROVIDER: S-EPMC6664181 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature

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The abundant DNA adduct <i>N</i><sup>7</sup>-methyl deoxyguanosine contributes to miscoding during replication by human DNA polymerase η.

Njuma Olive J OJ   Su Yan Y   Guengerich F Peter FP  

The Journal of biological chemistry 20190517 26


Aside from abasic sites and ribonucleotides, the DNA adduct <i>N</i><sup>7</sup>-methyl deoxyguanosine (<i>N<sup>7</sup></i> -CH<sub>3</sub> dG) is one of the most abundant lesions in mammalian DNA. Because <i>N<sup>7</sup></i> -CH<sub>3</sub> dG is unstable, leading to deglycosylation and ring-opening, its miscoding potential is not well-understood. Here, we employed a 2'-fluoro isostere approach to synthesize an oligonucleotide containing an analog of this lesion (<i>N<sup>7</sup></i> -CH<sub>  ...[more]

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