Research output: Contribution to journal › Article › peer-review
Characterization of DNA ADP-ribosyltransferase activities of PARP2 and PARP3 : New insights into DNA ADP-ribosylation. / Zarkovic, Gabriella; Belousova, Ekaterina A.; Talhaoui, Ibtissam et al.
In: Nucleic Acids Research, Vol. 46, No. 5, 16.03.2018, p. 2417-2431.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Characterization of DNA ADP-ribosyltransferase activities of PARP2 and PARP3
T2 - New insights into DNA ADP-ribosylation
AU - Zarkovic, Gabriella
AU - Belousova, Ekaterina A.
AU - Talhaoui, Ibtissam
AU - Saint-Pierre, Christine
AU - Kutuzov, Mikhail M.
AU - Matkarimov, Bakhyt T.
AU - Biard, Denis
AU - Gasparutto, Didier
AU - Lavrik, Olga I.
AU - Ishchenko, Alexander A.
N1 - Publisher Copyright: © The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.
PY - 2018/3/16
Y1 - 2018/3/16
N2 - 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.
AB - 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.
KW - Adenosine Diphosphate Ribose/metabolism
KW - Cell Cycle Proteins/metabolism
KW - DNA Adducts/metabolism
KW - DNA Breaks
KW - DNA Breaks, Double-Stranded
KW - DNA/chemistry
KW - HeLa Cells
KW - Humans
KW - Phosphates/metabolism
KW - Poly(ADP-ribose) Polymerases/metabolism
KW - Substrate Specificity
KW - PATHWAYS
KW - CELLS
KW - POLY(ADP-RIBOSE) POLYMERASE
KW - MOUSE
KW - TRANSCRIPTION ELONGATION
KW - TARGETS REVEALS
KW - DAMAGE REPAIR
KW - STRUCTURAL BASIS
KW - BREAKS
KW - DEPENDENT ACTIVATION
UR - http://www.scopus.com/inward/record.url?scp=85043399955&partnerID=8YFLogxK
U2 - 10.1093/nar/gkx1318
DO - 10.1093/nar/gkx1318
M3 - Article
C2 - 29361132
AN - SCOPUS:85043399955
VL - 46
SP - 2417
EP - 2431
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 5
ER -
ID: 16104692