Epigenetic Pyrimidine Nucleotides in Competition with Natural dNTPs as Substrates for Diverse DNA Polymerases

0558957 - ÚOCHB 2023 RIV US eng J - Journal Article
Pospíšil, Šimon - Panattoni, Alessandro - Gracias, Filip - Sýkorová, Veronika - Vaňková Hausnerová, Viola - Vítovská, Dragana - Šanderová, Hana - Krásný, Libor - Hocek, Michal
Epigenetic Pyrimidine Nucleotides in Competition with Natural dNTPs as Substrates for Diverse DNA Polymerases.
ACS Chemical Biology. Roč. 17, č. 10 (2022), s. 2781-2788. ISSN 1554-8929. E-ISSN 1554-8937
R&D Projects: GA ČR(CZ) GX20-00885X; GA MŠMT(CZ) EF16_019/0000729; GA ČR(CZ) GA22-12023S
Institutional support: RVO:61388963 ; RVO:61388971
Keywords : gene * cleavage * 5-hydroxymethyl
OECD category: Biochemistry and molecular biology
Impact factor: 4, year: 2022
Method of publishing: Open access
https://doi.org/10.1021/acschembio.2c00342

Five 2 '-deoxyribonucleoside triphosphates (dNTPs) derived from epigenetic pyrimidines (5-methylcytosine, 5hydroxymethylcytosine, 5-formylcytosine, 5-hydroxymethyluracil, and 5-formyluracil) were prepared and systematically studied as substrates for nine DNA polymerases in competition with natural dNTPs by primer extension experiments. The incorporation of these substrates was evaluated by a restriction endonucleases cleavage-based assay and by a kinetic study of single nucleotide extension. All of the modified pyrimidine dNTPs were good substrates for the studied DNA polymerases that incorporated a significant percentage of the modified nucleotides into DNA even in the presence of natural nucleotides. 5-Methylcytosine dNTP was an even better substrate for most polymerases than natural dCTP. On the other hand, 5-hydroxymethyl-2 '-deoxyuridine triphosphate was not the best substrate for SPO1 DNA polymerase, which naturally synthesizes 5hmU-rich genomes of the SPO1 bacteriophage. The results shed light onto the possibility of gene silencing through recycling and random incorporation of epigenetic nucleotides and into the replication of modified bacteriophage genomes.
Permanent Link: https://hdl.handle.net/11104/0332438