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 journalArticle

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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

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Poly(ADP-ribose) Polymerases
Adenosine Diphosphate Ribose
DNA
Phosphates
Adenosine Diphosphate
Ribose
Oligonucleotides
NAD
Deoxyribonucleotides
Poly Adenosine Diphosphate Ribose
Biological Phenomena
Single-Stranded DNA Breaks
Double-Stranded DNA Breaks
DNA Adducts
Glycoside Hydrolases
Nuclear Proteins
DNA Repair
Hydroxyl Radical
In Vitro Techniques
Mass Spectrometry

ASJC Scopus subject areas

  • Genetics

Cite this

Talhaoui, I., Lebedeva, N. A., Zarkovic, G., Saint-Pierre, C., Kutuzov, M. M., Sukhanova, M. V., ... Ishchenko, A. A. (2016). Poly(ADP-ribose) polymerases covalently modify strand break termini in DNA fragments in vitro. Nucleic Acids Research, 44(19), 9279-9295. https://doi.org/10.1093/nar/gkw675

Poly(ADP-ribose) polymerases covalently modify strand break termini in DNA fragments in vitro. / Talhaoui, Ibtissam; Lebedeva, Natalia A.; Zarkovic, Gabriella; Saint-Pierre, Christine; Kutuzov, Mikhail M.; Sukhanova, Maria V.; Matkarimov, Bakhyt T.; Gasparutto, Didier; Saparbaev, Murat K.; Lavrik, Olga I.; Ishchenko, Alexander A.

In: Nucleic Acids Research, Vol. 44, No. 19, 02.11.2016, p. 9279-9295.

Research output: Contribution to journalArticle

Talhaoui, I, Lebedeva, NA, Zarkovic, G, Saint-Pierre, C, Kutuzov, MM, Sukhanova, MV, Matkarimov, BT, Gasparutto, D, Saparbaev, MK, Lavrik, OI & Ishchenko, AA 2016, 'Poly(ADP-ribose) polymerases covalently modify strand break termini in DNA fragments in vitro', Nucleic Acids Research, vol. 44, no. 19, pp. 9279-9295. https://doi.org/10.1093/nar/gkw675
Talhaoui I, Lebedeva NA, Zarkovic G, Saint-Pierre C, Kutuzov MM, Sukhanova MV et al. Poly(ADP-ribose) polymerases covalently modify strand break termini in DNA fragments in vitro. Nucleic Acids Research. 2016 Nov 2;44(19):9279-9295. https://doi.org/10.1093/nar/gkw675
Talhaoui, Ibtissam ; Lebedeva, Natalia A. ; Zarkovic, Gabriella ; Saint-Pierre, Christine ; Kutuzov, Mikhail M. ; Sukhanova, Maria V. ; Matkarimov, Bakhyt T. ; Gasparutto, Didier ; Saparbaev, Murat K. ; Lavrik, Olga I. ; Ishchenko, Alexander A. / Poly(ADP-ribose) polymerases covalently modify strand break termini in DNA fragments in vitro. In: Nucleic Acids Research. 2016 ; Vol. 44, No. 19. pp. 9279-9295.
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