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

Gabriella Zarkovic (1) , Ekaterina A Belousova (2) , Ibtissam Talhaoui (3, 1) , Christine Saint-Pierre (4, 5) , Mikhail M Kutuzov (2) , Bakhyt T Matkarimov (6) , Denis Biard (7) , Didier Gasparutto (4, 5, 8) , Olga I Lavrik (2) , Alexander Ishchenko (3, 1)

1 UMR 8200 - Stabilité Génétique et Oncogenèse
2 ICBFM SB RAS - Institute of Chemical Biology and Fundamental Medicine [Novosibirsk, Russia]
3 IGC - Intégrité du génome et cancers
4 CREAB - Chimie pour la Reconnaissance et l’Etude d’Assemblages Biologiques
5 INAC - Institut Nanosciences et Cryogénie
6 National Laboratory Astana
7 JACOB - Institut de Biologie François JACOB
8 SYMMES - SYstèmes Moléculaires et nanoMatériaux pour l’Energie et la Santé

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. PARP3catalyzed 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, PARPmediated 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 PARP2catalyzed 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.

Domaines: Sciences du Vivant [q-bio]; Biologie moléculaire

Gabriella Zarkovic, Ekaterina A Belousova, Ibtissam Talhaoui, Christine Saint-Pierre, Mikhail M Kutuzov, et al.. Characterization of DNA ADP-ribosyltransferase activities of PARP2 and PARP3: new insights into DNA ADP-ribosylation. Nucleic Acids Research, 2018, 46 (5), pp.2417 - 2431. ⟨10.1093/nar/gkx1318⟩. ⟨hal-03423702⟩