Poly(ADP-ribose) polymerases covalently modify strand break termini in DNA fragments in vitro

Ibtissam Talhaoui, Natalia A. Lebedeva, Gabriella Zarkovic, Christine Saint-Pierre, Mikhail M. Kutuzov, Maria V. Sukhanova, Bakhyt T. Matkarimov, Didier Gasparutto, Murat K. Saparbaev, Olga I. Lavrik, Alexander A. Ishchenko

Research output: Contribution to journalArticlepeer-review

59 Citations (Scopus)

Abstract

Poly(ADP-ribose) polymerases (PARPs/ARTDs) use nicotinamide adenine dinucleotide (NAD+) to catalyse the synthesis of a long branched poly(ADP-ribose) polymer (PAR) attached to the acceptor amino acid residues of nuclear proteins. PARPs act on single- and double-stranded DNA breaks by recruiting DNA repair factors. Here, in in vitro biochemical experiments, we found that the mammalian PARP1 and PARP2 proteins can directly ADP-ribosylate the termini of DNA oligonucleotides. PARP1 preferentially catalysed covalent attachment of ADP-ribose units to the ends of recessed DNA duplexes containing 3′-cordycepin, 5′- and 3′-phosphate and also to 5′-phosphate of a single-stranded oligonucleotide. PARP2 preferentially ADP-ribosylated the nicked/gapped DNA duplexes containing 5′-phosphate at the double-stranded termini. PAR glycohydrolase (PARG) restored native DNA structure by hydrolysing PAR-DNA adducts generated by PARP1 and PARP2. Biochemical and mass spectrometry analyses of the adducts suggested that PARPs utilise DNA termini as an alternative to 2′-hydroxyl of ADP-ribose and protein acceptor residues to catalyse PAR chain initiation either via the 2′,1″-O-glycosidic ribose-ribose bond or via phosphodiester bond formation between C1′ of ADP-ribose and the phosphate of a terminal deoxyribonucleotide. This new type of post-replicative modification of DNA provides novel insights into the molecular mechanisms underlying biological phenomena of ADP-ribosylation mediated by PARPs.

Original languageEnglish
Pages (from-to)9279-9295
Number of pages17
JournalNucleic Acids Research
Volume44
Issue number19
DOIs
Publication statusPublished - Nov 2 2016

ASJC Scopus subject areas

  • Genetics

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