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Template Properties of 5-Methyl-2'-Deoxycytidine and 5-Hydroxymethyl-2'-Deoxycytidine in Reactions with Human Translesion and Reparative DNA Polymerases

  • ENZYMOLOGY OF DNA REPAIR SYSTEMS
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Abstract—

5-Methyl-2'-deoxycytidine (mC) and the product of its controlled oxidation, 5-hydroxymethyl-2'-cytidine (hmC), play a key role in the epigenetic regulation of gene expression, the cell differentiation, and the carcinogenesis. Due to spontaneious deamination, genomic CpG sites containing mC and hmC serve as mutagenesis hotspots. In addition, error-prone translesion and reparative DNA polymerases may serve as additional source of mutations in the lesion-containing regions with CpG sites. In the present work, we performed in vitro analysis of the accuracy of nucleotide incorporation opposite to mC and hmC by human DNA polymerases Polβ, Polλ, Polη, Polι, Polκ and primase polymerase PrimPol. The results of the study show a high accuracy of copying mC and hmC by the reparative DNA polymerases Polβ and Polλ, while Polη, Polι, Polκ, and PrimPol copied mC and hmC with less accuracy evident by incorporation of dAMP and dTMP. The same spectrum of error-prone dNMP incorporation was also noted at sites with unmodified cytosines.

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ACKNOWLEDGMENTS

We are grateful to L.V. Gening (Institute of Molecular Genetics) for the Pol κ preparation, O.I. Lavrik (Institute of Chemical Biology and Fundamental Medicine) for the Polλ expression vector, and K.A. Bondarenko for help in Polη purification.

Funding

This work was supported by the Russian Foundation for Basic Research (project nos. 17-00-00264-komfi (AVM) and 17-00-00261-komfi (DOZ)). PrimPol activity testing was supported by the Russian Science Foundation (project no. 18-14-00354 (AVM)).

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Authors and Affiliations

  1. Institute of Molecular Genetics, Kurchatov Institute Research Center, 123182, Moscow, Russia

    E. S. Shilkin, V. A. Poltorachenko, E. O. Boldinova & A. V. Makarova

  2. Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, 630090, Novosibirsk, Russia

    D. V. Petrova & D. O. Zharkov

  3. Novosibirsk State University, 630090, Novosibirsk, Russia

    D. O. Zharkov

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  1. E. S. Shilkin
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  2. D. V. Petrova
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  4. E. O. Boldinova
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  5. D. O. Zharkov
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  6. A. V. Makarova
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Correspondence to D. O. Zharkov or A. V. Makarova.

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The authors declare that they have no conflict of interest. This work does not contain any studies involving animals or human subjects performed by any of the authors.

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Translated by T. Tkacheva

Abbreviations: mC, 5-methyl-2'-deoxycytidine; hmC, 5-hydro-xymethyl-2'-deoxycytidine; oxoG, 8-oxo-7,8-dihydro-2'-deoxy-guanosine.

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Shilkin, E.S., Petrova, D.V., Poltorachenko, V.A. et al. Template Properties of 5-Methyl-2'-Deoxycytidine and 5-Hydroxymethyl-2'-Deoxycytidine in Reactions with Human Translesion and Reparative DNA Polymerases. Mol Biol 55, 267–272 (2021). https://doi.org/10.1134/S0026893321020138

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