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Repair of DNA damage induced by the mycotoxin alternariol involves tyrosyl-DNA phosphodiesterase 1

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Abstract

Alternariol (AOH) was reported recently to act as a topoisomerase poison. To underline the relevance of topoisomerase targeting for the genotoxic properties of AOH, we addressed the question whether human tyrosyl-DNA phosphodiesterase 1 (TDP1), an enzyme vital to the repair of covalent DNA-topoisomerase adducts, affects AOH-mediated genotoxicity. The relevance of TDP1 activity on AOH-induced genotoxicity was investigated by the comet assay in human cells overexpressing GFP chimera of TDP1 or the inactive mutant TDP1H263A as well as in cells subjected to siRNA-mediated knock-down of endogenous TDP1. Cells overexpressing TDP1 exhibited significantly less DNA damage after treatment with AOH in comparison to cells expressing the inactive mutant TDP1H263A. In accordance with these results, levels of AOH inducing DNA strand breaks were increased in TDP1-suppressed cells in comparison to cells transfected with control siRNA. The specific topoisomerase poisons camptothecin and etoposide caused comparable effects, underlining that TDP1 plays an important role in the repair of topoisomerase-mediated DNA damage. In summary, the repair enzyme TDP1 was identified as a factor for the modulation of AOH-mediated DNA damage in human cells.

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Acknowledgement

We gratefully acknowledge J. Podlech (Universität Karlsruhe (TH), Germany) for the synthesis of AOH and AME. The study was supported by the Karlsruher Zentrum für Lebensmittel und Gesundheit (KIT, Germany) and the Deutsche Forschungsgemeinschaft SFB 728.

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Authors and Affiliations

  1. Research Platform Molecular Food Science, Department of Food Chemistry and Toxicology, University of Vienna, Waehringer Str. 38, 1090, Vienna, Austria

    Simone Baechler, Christopher Kropat, Gudrun Pahlke & Doris Marko

  2. Institute of Applied Biosciences, Section of Food Toxicology, Universität Karlsruhe (TH), Adenauerring 20, 76131, Karlsruhe, Germany

    Markus Fehr

  3. Medical School, Institute of Clinical Chemistry and Laboratory Diagnostics, Heinrich-Heine-University, 40225, Düsseldorf, Germany

    Christian Mielke & Fritz Boege

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  1. Markus Fehr
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  2. Simone Baechler
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  3. Christopher Kropat
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  4. Christian Mielke
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  5. Fritz Boege
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  6. Gudrun Pahlke
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  7. Doris Marko
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Correspondence to Doris Marko.

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Fehr, M., Baechler, S., Kropat, C. et al. Repair of DNA damage induced by the mycotoxin alternariol involves tyrosyl-DNA phosphodiesterase 1. Mycotox Res 26, 247–256 (2010). https://doi.org/10.1007/s12550-010-0063-6

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  • DOI: https://doi.org/10.1007/s12550-010-0063-6

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