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Vol 55(2021) N 2 p. 269-274; DOI 10.1134/S0026893321020138 Full Text

E.S. Shilkin1, D.V. Petrova2, V.A. Poltorachenko1, E.O. Boldinova1, D.O. Zharkov2,3*, A.V. Makarova1**

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

1Institute of Molecular Genetics, Kurchatov Institute Research Center, Moscow, 123182 Russia
2Institute of Chemical Biology and Fundamental Medicine, Siberian Branch, Russian Academy of Sciences, Novosibirsk, 630090 Russia
3Novosibirsk State University, Novosibirsk, 630090 Russia

*dzharkov@niboch.nsc.ru
**amakarova-img@yandex.ru
Received - 2020-08-30; Revised - 2020-09-18; Accepted - 2020-09-24

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, PoIκ and primase polymerase PrimPol. The results of the study show a high accuracy of copying mC and hmC by the reparative DNA polymerases polymerases Polβ and Polλ, while Polη, Polι, PoIκ, 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.

CpG sites, 5-methylcytosine, 5-hydroxymethylcytosine, translesion DNA synthesis, mutagenesis



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