Repair and DNA Polymerase Bypass of Clickable Pyrimidine Nucleotides | Sciact - CRIS-система Института катализа

Repair and DNA Polymerase Bypass of Clickable Pyrimidine Nucleotides Научная публикация

Журнал

Biomolecules
ISSN: 2218-273X

Вых. Данные

Год: 2024, Том: 14, Номер: 6, Номер статьи : 681, Страниц : 16 DOI: 10.3390/biom14060681

Ключевые слова

click chemistry; metabolic labeling; DNA repair; replication; DNA glycosylases; DNA polymerases; 5-ethynyl-2′-deoxyuridine

Авторы

Endutkin Anton V. ¹ , Yudkina Anna V. ¹ , Zharkov Timofey D. ¹ , Barmatov Alexander E. ¹ , Petrova Daria V. ¹ , Kim Daria V. ¹ , Zharkov Dmitry O. ^2,1

Организации

1	Siberian Branch of the Russian Academy of Sciences Institute of Chemical Biology and Fundamental Medicine, 8 Lavrentieva Ave., 630090 Novosibirsk, Russia
2	Department of Natural Sciences, Novosibirsk State University, 2 Pirogova St., 630090 Novosibirsk, Russia

Clickable nucleosides, most often 5-ethynyl-2′-deoxyuridine (EtU), are widely used in studies of DNA replication in living cells and in DNA functionalization for bionanotechology applications. Although clickable dNTPs are easily incorporated by DNA polymerases into the growing chain, afterwards they might become targets for DNA repair systems or interfere with faithful nucleotide insertion. Little is known about the possibility and mechanisms of these post-synthetic events. Here, we investigated the repair and (mis)coding properties of EtU and two bulkier clickable pyrimidine nucleosides, 5-(octa-1,7-diyn-1-yl)-U (C8-AlkU) and 5-(octa-1,7-diyn-1-yl)-C (C8-AlkC). In vitro, EtU and C8-AlkU, but not C8-AlkC, were excised by SMUG1 and MBD4, two DNA glycosylases from the base excision repair pathway. However, when placed into a plasmid encoding a fluorescent reporter inactivated by repair in human cells, EtU and C8-AlkU persisted for much longer than uracil or its poorly repairable phosphorothioate-flanked derivative. DNA polymerases from four different structural families preferentially bypassed EtU, C8-AlkU and C8-AlkC in an error-free manner, but a certain degree of misincorporation was also observed, especially evident for DNA polymerase β. Overall, clickable pyrimidine nucleotides could undergo repair and be a source of mutations, but the frequency of such events in the cell is unlikely to be considerable.

Endutkin A.V. , Yudkina A.V. , Zharkov T.D. , Barmatov A.E. , Petrova D.V. , Kim D.V. , Zharkov D.O.
Repair and DNA Polymerase Bypass of Clickable Pyrimidine Nucleotides
Biomolecules. 2024. V.14. N6. 681 :1-16. DOI: 10.3390/biom14060681 WOS Scopus OpenAlex

Поступила в редакцию:	12 мая 2024 г.
Принята к публикации:	10 июн. 2024 г.
Опубликована в печати:	12 июн. 2024 г.
Опубликована online:	12 июн. 2024 г.

Web of science:	WOS:001254583000001
Scopus:	2-s2.0-85197126697
OpenAlex:	W4399576585

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