title: Poly(ADP-ribose) polymerases covalently modify strand break termini in DNA fragments in vitro.
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.
authors.affiliation: Laboratoire «Stabilité Génétique et Oncogenèse» CNRS, UMR 8200, Univ. Paris-Sud, Université Paris-Saclay, F-94805 Villejuif, France.|Gustave Roussy, Université Paris-Saclay, F-94805 Villejuif, France.
authors.forename: Ibtissam
authors.initials: I
authors.lastname: Talhaoui
authors.affiliation: SB RAS Institute of Chemical Biology and Fundamental Medicine, Lavrentiev Av. 8, Novosibirsk 630090, Russia.
authors.forename: Natalia A
authors.initials: NA
authors.lastname: Lebedeva
authors.affiliation: Laboratoire «Stabilité Génétique et Oncogenèse» CNRS, UMR 8200, Univ. Paris-Sud, Université Paris-Saclay, F-94805 Villejuif, France.|Gustave Roussy, Université Paris-Saclay, F-94805 Villejuif, France.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Zarkovic
authors.affiliation: Université Grenoble Alpes, CEA, INAC/SPrAM UMR5819 CEA CNRS UGA, F-38000 Grenoble, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Saint-Pierre
authors.affiliation: SB RAS Institute of Chemical Biology and Fundamental Medicine, Lavrentiev Av. 8, Novosibirsk 630090, Russia.
authors.forename: Mikhail M
authors.initials: MM
authors.lastname: Kutuzov
authors.affiliation: SB RAS Institute of Chemical Biology and Fundamental Medicine, Lavrentiev Av. 8, Novosibirsk 630090, Russia.
authors.forename: Maria V
authors.initials: MV
authors.lastname: Sukhanova
authors.affiliation: National Laboratory Astana, Nazarbayev University, Astana 010000, Kazakhstan.
authors.forename: Bakhyt T
authors.initials: BT
authors.lastname: Matkarimov
authors.affiliation: Université Grenoble Alpes, CEA, INAC/SPrAM UMR5819 CEA CNRS UGA, F-38000 Grenoble, France.
authors.forename: Didier
authors.initials: D
authors.lastname: Gasparutto
authors.affiliation: Laboratoire «Stabilité Génétique et Oncogenèse» CNRS, UMR 8200, Univ. Paris-Sud, Université Paris-Saclay, F-94805 Villejuif, France.|Gustave Roussy, Université Paris-Saclay, F-94805 Villejuif, France.
authors.forename: Murat K
authors.initials: MK
authors.lastname: Saparbaev
authors.affiliation: SB RAS Institute of Chemical Biology and Fundamental Medicine, Lavrentiev Av. 8, Novosibirsk 630090, Russia lavrik@niboch.nsc.ru.|Department of Natural Sciences, Novosibirsk State University, 2 Pirogova St., Novosibirsk 630090, Russia.
authors.forename: Olga I
authors.initials: OI
authors.lastname: Lavrik
authors.affiliation: Laboratoire «Stabilité Génétique et Oncogenèse» CNRS, UMR 8200, Univ. Paris-Sud, Université Paris-Saclay, F-94805 Villejuif, France Alexander.Ishchenko@gustaveroussy.fr.|Gustave Roussy, Université Paris-Saclay, F-94805 Villejuif, France.
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Ishchenko
mesh_terms: D000818:Animals; D002384:Catalysis; D004247:DNA / Q000235:genetics* / Q000378:metabolism*; D018736:DNA Adducts; D053903:DNA Breaks, Double-Stranded; D006801:Humans; D006868:Hydrolysis; D051379:Mice; D009243:NAD / Q000378:metabolism; D000071137:Poly (ADP-Ribose) Polymerase-1 / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D011485:Protein Binding; D013379:Substrate Specificity
chemical_list: D018736:DNA Adducts; D009243:NAD; D004247:DNA; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; C459820:Parp2 protein, mouse