Research Paper Volume 11, Issue 9 pp 2852—2873
Poly(ADP-ribosyl)ation and DNA repair synthesis in the extracts of naked mole rat, mouse, and human cells
- 1 Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk 630090, Russia
- 2 Novosibirsk State University, Novosibirsk 630090, Russia
- 3 Institute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of Sciences, Novosibirsk 630090, Russia
Received: March 6, 2019 Accepted: May 3, 2019 Published: May 13, 2019
https://doi.org/10.18632/aging.101959How to Cite
Copyright: Kosova et al. This is an open‐access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
DNA repair capacity in cells of naked mole rat (Hgl), a species known for its longevity and resistance to cancer, is still poorly characterized. Here, using the whole-cell extracts (WCEs) of Hgl, mouse and human cells, we studied the interrelation between DNA synthesis on the substrates of base excision repair and the activity of poly(ADP-ribose) polymerases (PARPs) responsible for the transfer of the ADP-ribose moieties onto different targets. The level of PAR synthesis was more than ten-fold higher in human WCE as compared to rodent WCEs, while the efficiency of DNA synthesis was comparable. Under conditions of PAR synthesis, the efficiency of DNA synthesis was only slightly enhanced in all extracts and in mouse WCEs unusual products of the primer elongation were detected. The results obtained with WCEs, recombinant proteins and recently found ability of PARPs to attach the ADP-ribose moieties to DNA allowed us to attribute these products to primer mono(ADP-ribosyl)ation (MARylation) at the 5ʹ-terminal phosphate by PARP3 during the DNA synthesis. PARP1/PARP2 can then transfer the ADP-ribose moieties onto initial ADP-ribose. Our results suggest that MARylation/PARylation of DNA in the extracts depends on the ratios between PARPs and can be controlled by DNA-binding proteins.
Abbreviations
AP site: apurinic/apyrimidinic (abasic) site; APE1: apurinic/apyrimidinic endonuclease 1; BER: base excision repair; DDR: DNA damage response; dRP: deoxyribose phosphate; DSB: double-strand break; FAP-dCTP: exo-N-{2-[N-(4-azido-2,5-difluoro-3-chloropyridine-6-yl)-3-aminopropionyl]aminoethyl}-2′-deoxycitidine-5′-triphosphate; Hgl: naked mole rat (Heterocephalus glaber); LP: long-patch; MARylation: mono(ADP-ribosyl)ation; Mmu: mouse (Mus musculus); NAD+: nicotinamide adenine dinucleotide; NER: nucleotide excision repair; PAGE: polyacrylamide gel electrophoresis; PAR: polymer of ADP-ribose; PARG: poly(ADP-ribose) glycohydrolase; PARP1/2/3: poly(ADP-ribose) polymerase 1/2/3; PARylation: poly(ADP-ribosyl)ation; рDEG: 5ʹ diethylene glycol phosphate; Polβ: DNA polymerase β; SP: short-patch; SSB: single-strand break; Ung: E. coli uracil-DNA glycosylase; WCE: whole-cell extract.