Interaction of PARP-2 with AP site containing DNA.
In eukaryotes the stability of genome is provided by functioning of DNA repair systems. One of the main DNA repair pathways in eukaryotes is the base excision repair (BER). This system requires precise regulation for correct functioning. Two members of the PARP family - PARP-1 and PARP-2, which can be activated by DNA damage - are widely considered as regulators of DNA repair processes, including BER. In contrast to PARP-1, the role of PARP-2 in BER has not been extensively studied yet. Since AP site is one of the most frequent type of DNA damage and a key intermediate of BER at the stage preceding formation of DNA breaks, in this paper we focused on the characterization of PARP-2 interaction with AP site-containing DNAs. We demonstrated that PARP-2, like PARP-1, can interact with the intact AP site via Schiff base formation, in spite of crucial difference in the structure of the DNA binding domains of these PARPs. By cross-linking of PARPs to AP DNA, we determined that the N-terminal domains of both PARPs are involved in formation of cross-links with AP DNA. We have also confirmed that DNA binding by PARP-2, in contrast to PARP-1, is not modulated by autoPARylation. PARP-2, like PARP-1, can inhibit the activity of APE1 by binding to AP site, but, in contrast to PARP-1, this inhibitory influence is hardly regulated by PAR synthesis. At the same time, 5'-dRP lyase activity of both PARPs is comparable, although being much weaker than that of Pol β, which is considered as the main 5'-dRP lyase of the BER process.
Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia.
Mikhail M
MM
Kutuzov
Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia; Novosibirsk State University, Novosibirsk, Russia.
Svetlana N
SN
Khodyreva
Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia; Novosibirsk State University, Novosibirsk, Russia.
Ekaterina S
ES
Ilina
Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia.
Maria V
MV
Sukhanova
UMR7242, Biotechnology and Cell Signaling, Université de Strasbourg, CNRS, MEDALIS, ESBS, Illkirch, France.
Jean-Christophe
JC
Amé
Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia; Novosibirsk State University, Novosibirsk, Russia. Electronic address: lavrik@niboch.nsc.ru.
Olga I
OI
Lavrik
D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D020218:Response Elements
D004247:DNA; C499320:PARP1 protein, human; C483041:PARP2 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; C085108:APEX1 protein, human; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase
AP lyase; AP sites; DNA repair; PARP-2; Schiff base