Abstract
The 1,N6-ethenodeoxyadenosine (εdA) lesion is promutagenic and has been implicated in carcinogenesis. We show here that human Polι, a Y-family DNA polymerase, can promote replication through this lesion by proficiently incorporating a nucleotide opposite it. The structural basis of this action is rotation of the εdA adduct to the syn conformation in the Polι active site and presentation of its 'Hoogsteen edge' for hydrogen-bonding with incoming dTTP or dCTP. We also show that Polζ carries out the subsequent extension reaction and that efficiency of extension from εdA·T is notably higher than from εdA·C. Together, our studies reveal for the first time how the exocyclic εdA adduct is accommodated in a DNA polymerase active site, and they show that the combined action of Polι and Polζ provides for efficient and error-free synthesis through this potentially carcinogenic DNA lesion.
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Acknowledgements
We thank the staff at the Advanced Photon Source (beamline 19ID) and Brookhaven National Laboratory (beamline X29) for facilitating X-ray data collection. We thank S. Townson and S. Lone for help with data collection and DNA purification. This work was supported by grant CA115856 from the US National Institutes of Health (S.P. and A.K.A.).
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D.T.N. performed the crystallographic experiments, R.E.J. conducted the kinetic experiments and all of the authors contributed to the concepts and to the writing of the paper.
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Nair, D., Johnson, R., Prakash, L. et al. Hoogsteen base pair formation promotes synthesis opposite the 1,N6-ethenodeoxyadenosine lesion by human DNA polymerase ι. Nat Struct Mol Biol 13, 619–625 (2006). https://doi.org/10.1038/nsmb1118
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DOI: https://doi.org/10.1038/nsmb1118
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