title: DNA damage and repair kinetics of the Alternaria mycotoxins alternariol, altertoxin II and stemphyltoxin III in cultured cells.
abstract: The Alternaria mycotoxins alternariol (AOH) and altertoxin II (ATX II) have previously been shown to elicit mutagenic and genotoxic effects in bacterial and mammalian cells, although with vastly different activities. For example, ATX II was about 50 times more mutagenic than AOH. We now report that stemphyltoxin III (STTX III) is also highly mutagenic. The more pronounced effects of the perylene quinones ATX II and STTX III at lower concentrations compared to the dibenzo-α-pyrone AOH indicate a marked dependence of the genotoxic potential on the chemical structure and furthermore suggest that the underlying modes of action may be different. We have now further investigated the type of DNA damage induced by AOH, ATX II and STTX III, as well as the repair kinetics and their dependence on the status of nucleotide excision repair (NER). DNA double strand breaks induced by AOH due to poisoning of topoisomerase IIα were completely repaired in less than 2h. Under cell-free conditions, inhibition of topoisomerase IIα could also be measured for ATX II and STTX III at low concentrations, but the perylene quinones were catalytic inhibitors rather than topoisomerase poisons and did not induce DSBs. DNA strand breaks induced by ATX II and STTX III were more persistent and not completely repaired within 24h. A dependence of the repair rate on the NER status could only be demonstrated for STTX III, resulting in an accumulation of DNA damage in NER-deficient cells. Together with the finding that the DNA glycosylase formamidopyrimidine-DNA glycosylase (Fpg), but not T4 endonuclease V, is able to generate additional DNA strand breaks measurable by the alkaline unwinding assay, we conclude that the genotoxicity of the perylene quinones with an epoxide group is probably caused by the formation of DNA adducts which may be converted to Fpg sensitive sites.
authors.affiliation: Institute of Applied Biosciences, Department of Food Chemistry and Toxicology, Karlsruhe Institute of Technology (KIT), Adenauerring 20a, D-76131 Karlsruhe, Germany.
authors.forename: Stefanie C
authors.initials: SC
authors.lastname: Fleck
authors.affiliation: Institute of Applied Biosciences, Department of Food Chemistry and Toxicology, Karlsruhe Institute of Technology (KIT), Adenauerring 20a, D-76131 Karlsruhe, Germany.
authors.forename: Friederike
authors.initials: F
authors.lastname: Sauter
authors.affiliation: Institute of Applied Biosciences, Department of Food Chemistry and Toxicology, Karlsruhe Institute of Technology (KIT), Adenauerring 20a, D-76131 Karlsruhe, Germany.
authors.forename: Erika
authors.initials: E
authors.lastname: Pfeiffer
authors.affiliation: Institute of Applied Biosciences, Department of Food Chemistry and Toxicology, Karlsruhe Institute of Technology (KIT), Adenauerring 20a, D-76131 Karlsruhe, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Metzler
authors.affiliation: Institute of Applied Biosciences, Department of Food Chemistry and Toxicology, Karlsruhe Institute of Technology (KIT), Adenauerring 20a, D-76131 Karlsruhe, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Hartwig
authors.affiliation: Institute of Applied Biosciences, Department of Food Chemistry and Toxicology, Karlsruhe Institute of Technology (KIT), Adenauerring 20a, D-76131 Karlsruhe, Germany. Electronic address: beate.koeberle@kit.edu.
authors.forename: Beate
authors.initials: B
authors.lastname: Köberle
mesh_terms: D000528:Alternaria; D000951:Antigens, Neoplasm / Q000378:metabolism; D001551:Benz(a)Anthracenes / Q000633:toxicity*; D002460:Cell Line; D004249:DNA Damage; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D006801:Humans; D007783:Lactones / Q000633:toxicity*; D009152:Mutagenicity Tests; D009153:Mutagens / Q000633:toxicity*; D009183:Mycotoxins / Q000633:toxicity*; D010569:Perylene / Q000031:analogs & derivatives* / Q000633:toxicity
chemical_list: D000951:Antigens, Neoplasm; D001551:Benz(a)Anthracenes; D004268:DNA-Binding Proteins; D007783:Lactones; D009153:Mutagens; D009183:Mycotoxins; C060436:stemphyltoxin III; C050234:altertoxin II; D010569:Perylene; D004250:DNA Topoisomerases, Type II; C005197:alternariol
keywords: Alternariol; Altertoxin II; DNA strand breaks; Mutagenicity; Nucleotide excision repair; Stemphyltoxin III