Homologues of epigenetic pyrimidines: 5-alkyl-, 5-hydroxyalkyl and 5-acyluracil andcytosine nucleotides: synthesis, enzymatic incorporation into DNA and effect on transcription with bacterial RNA polymerase

0559506 - ÚOCHB 2023 RIV GB eng J - Journal Article
Gracias, Filip - Ruiz-Larrabeiti, Olatz - Vaňková Hausnerová, Viola - Pohl, Radek - Klepetářová, Blanka - Sýkorová, Veronika - Krásný, Libor - Hocek, Michal
Homologues of epigenetic pyrimidines: 5-alkyl-, 5-hydroxyalkyl and 5-acyluracil andcytosine nucleotides: synthesis, enzymatic incorporation into DNA and effect on transcription with bacterial RNA polymerase.
RSC Chemical Biology. Roč. 3, č. 8 (2022), s. 1069-1075. ISSN 2633-0679. E-ISSN 2633-0679
R&D Projects: GA MŠMT(CZ) EF16_019/0000729; GA ČR(CZ) GX20-00885X; GA ČR(CZ) GA22-12023S
Institutional support: RVO:61388963 ; RVO:61388971
Keywords : major-groove * 5-formylcytosine * 5-hydroxymethylcytosine
OECD category: Biochemistry and molecular biology; Microbiology (MBU-M)
Impact factor: 4.1, year: 2022
Method of publishing: Open access
https://doi.org/10.1039/D2CB00133K

Homologues of natural epigenetic pyrimidine nucleosides and nucleotides were designed and synthesized. They included 5-ethyl-, 5-propyl-, 5-(1-hydroxyethyl)-, 5-(1-hydroxypropyl)- and 5-acetyl- and 5-propionyl-cytosine anduracil 2'-deoxyribonucleosides and their corresponding 5'-O-triphosphates (dN(x)TPs). The epimers of 5-(1-hydroxyethyl)- and 5-(1-hydroxypropyl)pyrimidine nucleosides were separated and their absolute configuration was determined by a combination of X-ray and NMR analysis. The modified dN(x)TPs were used as substrates for PCR synthesis of modified DNA templates used for the study of transcription with bacterial RNA polymerase. Fundamental differences in transcription efficiency were observed, depending on the various modifications. The most notable effects included pronounced stimulation of transcription from 5-ethyluracil-bearing templates (200% transcription yield compared to natural thymine) and an enhancing effect of 5-acetylcytosine versus inhibiting effect of 5-acetyluracil. In summary, these results reveal that RNA polymerase copes with dramatically altered DNA structure and suggest that these nucleobases could potentially play roles as artificial epigenetic DNA nucleobases.
Permanent Link: https://hdl.handle.net/11104/0332783


Research data: CCDC, CCDC