title: Lipid peroxidation product 4-hydroxy-2-nonenal modulates base excision repair in human cells.
abstract: Oxidative-stress-driven lipid peroxidation (LPO) is involved in the pathogenesis of several human diseases, including cancer. LPO products react with cellular proteins changing their properties, and with DNA bases to form mutagenic etheno-DNA adducts, removed from DNA mainly by the base excision repair (BER) pathway. One of the major reactive aldehydes generated by LPO is 4-hydroxy-2-nonenal (HNE). We investigated the effect of HNE on BER enzymes in human cells and in vitro. K21 cells pretreated with physiological HNE concentrations were more sensitive to oxidative and alkylating agents, H2O2 and MMS, than were untreated cells. Detailed examination of the effects of HNE on particular stages of BER in K21 cells revealed that HNE decreases the rate of excision of 1,N(6)-ethenoadenine (ɛA) and 3,N(4)-ethenocytosine (ɛC), but not of 8-oxoguanine. Simultaneously HNE increased the rate of AP-site incision and blocked the re-ligation step after the gap-filling by DNA polymerases. This suggested that HNE increases the number of unrepaired single-strand breaks (SSBs) in cells treated with oxidizing or methylating agents. Indeed, preincubation of cells with HNE and their subsequent treatment with H2O2 or MMS increased the number of nuclear poly(ADP-ribose) foci, known to appear in cells in response to SSBs. However, when purified BER enzymes were exposed to HNE, only ANPG and TDG glycosylases excising ɛA and ɛC from DNA were inhibited, and only at high HNE concentrations. APE1 endonuclease and 8-oxoG-DNA glycosylase 1 (OGG1) were not inhibited. These results indicate that LPO products exert their promutagenic action not only by forming DNA adducts, but in part also by compromising the BER pathway.
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland.
authors.forename: Alicja
authors.initials: A
authors.lastname: Winczura
authors.affiliation: Institute of Biochemistry, Faculty of Biology, University of Warsaw, Miecznikowa 1, Warsaw, Poland.
authors.forename: Alicja
authors.initials: A
authors.lastname: Czubaty
authors.affiliation: Institute of Biochemistry, Faculty of Biology, University of Warsaw, Miecznikowa 1, Warsaw, Poland.
authors.forename: Kinga
authors.initials: K
authors.lastname: Winczura
authors.affiliation: Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw, Pawińskiego 5a, Warsaw, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Masłowska
authors.affiliation: Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw, Pawińskiego 5a, Warsaw, Poland.
authors.forename: Matylda
authors.initials: M
authors.lastname: Nałęcz
authors.affiliation: Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw, Pawińskiego 5a, Warsaw, Poland.
authors.forename: Dominika A
authors.initials: DA
authors.lastname: Dudzińska
authors.affiliation: Groupe «Réparation de l'ADN», CNRS UMR8200, Université Paris-Sud, Institut de Cancérologie Gustave Roussy, F-94805 Villejuif Cedex, France.
authors.forename: Murat
authors.initials: M
authors.lastname: Saparbaev
authors.affiliation: Institute of Biochemistry, Faculty of Biology, University of Warsaw, Miecznikowa 1, Warsaw, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Staroń
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland; Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw, Pawińskiego 5a, Warsaw, Poland. Electronic address: tudek@ibb.waw.pl.
authors.forename: Barbara
authors.initials: B
authors.lastname: Tudek
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000378:metabolism; D000447:Aldehydes / Q000378:metabolism / Q000494:pharmacology*; D002460:Cell Line; D003596:Cytosine / Q000031:analogs & derivatives / Q000378:metabolism; D053904:DNA Breaks, Single-Stranded; D045647:DNA Glycosylases / Q000037:antagonists & inhibitors / Q000378:metabolism; D004260:DNA Repair / Q000187:drug effects*; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000378:metabolism; D006147:Guanine / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D015227:Lipid Peroxidation
chemical_list: C018289:3,N(4)-ethenocytosine; D000447:Aldehydes; C004981:1,N(6)-ethenoadenine; C024829:8-hydroxyguanine; D006147:Guanine; D003596:Cytosine; D045647:DNA Glycosylases; C479058:oxoguanine glycosylase 1, human; C085108:APEX1 protein, human; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D000225:Adenine; C027576:4-hydroxy-2-nonenal
keywords: 4-Hydroxy-2-nonenal; Base excision repair; HNE–DNA adducts; Ligation, OGG1, TDG, ANPG, APE1