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Article Dans Une Revue Nucleic Acids Research Année : 2016

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

Résumé

Poly(ADP-ribose) polymerases (PARPs/ARTDs) use nicotinamide adenine dinucleotide (NAD +) to catalyse the synthesis of a long branched poly(ADPribose) 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 3phosphate and also to 5-phosphate of a singlestranded oligonucleotide. PARP2 preferentially ADPribosylated 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 ADPribose 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.
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Dates et versions

hal-03419285 , version 1 (08-11-2021)

Identifiants

Citer

Ibtissam Talhaoui, Natalia A Lebedeva, Gabriella Zarkovic, Christine Saint-Pierre, Mikhail M Kutuzov, et al.. Poly(ADP-ribose) polymerases covalently modify strand break termini in DNA fragments in vitro. Nucleic Acids Research, 2016, 44 (19), pp.9279-9295. ⟨10.1093/nar/gkw675⟩. ⟨hal-03419285⟩
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