Characterization of DNA ADP-ribosyltransferase activities of PARP2 and PARP3: New insights into DNA ADP-ribosylation

Gabriella Zarkovic, Ekaterina A. Belousova, Ibtissam Talhaoui, Christine Saint-Pierre, Mikhail M. Kutuzov, Bakhyt Matkarimov, Denis Biard, Didier Gasparutto, Olga I. Lavrik, Alexander A. Ishchenko

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Abstract

Poly(ADP-ribose) polymerases (PARPs) act as DNA break sensors and catalyze the synthesis of polymers of ADP-ribose (PAR) covalently attached to acceptor proteins at DNA damage sites. It has been demonstrated that both mammalian PARP1 and PARP2 PARylate double-strand break termini in DNA oligonucleotide duplexes in vitro. Here, we show that mammalian PARP2 and PARP3 can PARylate and mono(ADP-ribosyl)ate (MARylate), respectively, 5- and 3-terminal phosphate residues at double- and single-strand break termini of a DNA molecule containing multiple strand breaks. PARP3-catalyzed DNA MARylation can be considered a new type of reversible post-replicative DNA modification. According to DNA substrate specificity of PARP3 and PARP2, we propose a putative mechanistic model of PARP-catalyzed strand break-oriented ADP-ribosylation of DNA termini. Notably, PARP-mediated DNA ADP-ribosylation can be more effective than PARPs' auto-ADP-ribosylation depending on the DNA substrates and reaction conditions used. Finally, we show an effective PARP3- or PARP2-catalyzed ADP-ribosylation of high-molecular-weight (∼3-kb) DNA molecules, PARP-mediated DNA PARylation in cell-free extracts and a persisting signal of anti-PAR antibodies in a serially purified genomic DNA from bleomycin-treated poly(ADP-ribose) glycohydrolase-depleted HeLa cells. These results suggest that certain types of complex DNA breaks can be effectively ADP-ribosylated by PARPs in cellular response to DNA damage.

Original languageEnglish
Pages (from-to)2417-2431
Number of pages15
JournalNucleic Acids Research
Volume46
Issue number5
DOIs
Publication statusPublished - Jan 1 2018

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ADP Ribose Transferases
Adenosine Diphosphate
Poly(ADP-ribose) Polymerases
DNA
Adenosine Diphosphate Ribose
DNA Breaks
DNA Damage
Polymers
Bleomycin
Substrate Specificity
Cell Extracts
HeLa Cells
Oligonucleotides

ASJC Scopus subject areas

  • Genetics

Cite this

Zarkovic, G., Belousova, E. A., Talhaoui, I., Saint-Pierre, C., Kutuzov, M. M., Matkarimov, B., ... Ishchenko, A. A. (2018). Characterization of DNA ADP-ribosyltransferase activities of PARP2 and PARP3: New insights into DNA ADP-ribosylation. Nucleic Acids Research, 46(5), 2417-2431. https://doi.org/10.1093/nar/gkx1318

Characterization of DNA ADP-ribosyltransferase activities of PARP2 and PARP3 : New insights into DNA ADP-ribosylation. / Zarkovic, Gabriella; Belousova, Ekaterina A.; Talhaoui, Ibtissam; Saint-Pierre, Christine; Kutuzov, Mikhail M.; Matkarimov, Bakhyt; Biard, Denis; Gasparutto, Didier; Lavrik, Olga I.; Ishchenko, Alexander A.

In: Nucleic Acids Research, Vol. 46, No. 5, 01.01.2018, p. 2417-2431.

Research output: Contribution to journalArticle

Zarkovic, G, Belousova, EA, Talhaoui, I, Saint-Pierre, C, Kutuzov, MM, Matkarimov, B, Biard, D, Gasparutto, D, Lavrik, OI & Ishchenko, AA 2018, 'Characterization of DNA ADP-ribosyltransferase activities of PARP2 and PARP3: New insights into DNA ADP-ribosylation', Nucleic Acids Research, vol. 46, no. 5, pp. 2417-2431. https://doi.org/10.1093/nar/gkx1318
Zarkovic, Gabriella ; Belousova, Ekaterina A. ; Talhaoui, Ibtissam ; Saint-Pierre, Christine ; Kutuzov, Mikhail M. ; Matkarimov, Bakhyt ; Biard, Denis ; Gasparutto, Didier ; Lavrik, Olga I. ; Ishchenko, Alexander A. / Characterization of DNA ADP-ribosyltransferase activities of PARP2 and PARP3 : New insights into DNA ADP-ribosylation. In: Nucleic Acids Research. 2018 ; Vol. 46, No. 5. pp. 2417-2431.
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