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 T. Matkarimov, Denis Biard, Didier Gasparutto, Olga I. Lavrik, Alexander A. Ishchenko

Research output: Contribution to journalArticlepeer-review

57 Citations (Scopus)

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 - 16 Mar 2018

Keywords

  • Adenosine Diphosphate Ribose/metabolism
  • Cell Cycle Proteins/metabolism
  • DNA Adducts/metabolism
  • DNA Breaks
  • DNA Breaks, Double-Stranded
  • DNA/chemistry
  • HeLa Cells
  • Humans
  • Phosphates/metabolism
  • Poly(ADP-ribose) Polymerases/metabolism
  • Substrate Specificity
  • PATHWAYS
  • CELLS
  • POLY(ADP-RIBOSE) POLYMERASE
  • MOUSE
  • TRANSCRIPTION ELONGATION
  • TARGETS REVEALS
  • DAMAGE REPAIR
  • STRUCTURAL BASIS
  • BREAKS
  • DEPENDENT ACTIVATION

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