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DNA repair defects sensitize cells to anticodon nuclease yeast killer toxins

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Abstract

Killer toxins from Kluyveromyces lactis (zymocin) and Pichia acaciae (PaT) were found to disable translation in target cells by virtue of anticodon nuclease (ACNase) activities on tRNAGlu and tRNAGln, respectively. Surprisingly, however, ACNase exposure does not only impair translation, but also affects genome integrity and concomitantly DNA damage occurs. Previously, it was shown that homologous recombination protects cells from ACNase toxicity. Here, we have analyzed whether other DNA repair pathways are functional in conferring ACNase resistance as well. In addition to HR, base excision repair (BER) and postreplication repair (PRR) promote clear resistance to either, PaT and zymocin. Comparative toxin sensitivity analysis of BER mutants revealed that its ACNase protective function is due to the endonucleases acting on apurinic (AP) sites, whereas none of the known DNA glycosylases is involved. Because PaT and zymocin require the presence of the ELP3/TRM9-dependent wobble uridine modification 5-methoxy-carbonyl-methyl (mcm5) for tRNA cleavage, we analyzed toxin response in DNA repair mutants additionally lacking such tRNA modifications. ACNase resistance caused by elp3 or trm9 mutations was found to rescue hypersensitivity of DNA repair defects, consistent with DNA damage to occur as a consequence of tRNA cleavage. The obtained genetic evidence promises to reveal new aspects into the mechanism linking translational fidelity and genome surveillance.

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Acknowledgments

Thanks are due to Dr. W. Kramer (Göttingen, Germany) for providing yeast strains. Financial support by the Deutsche Forschungsgemeinschaft (DFG) Grant no. ME 1142/5-1 is gratefully acknowledged.

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Author notes

  1. Roland Klassen

    Present address: Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, 77555, USA

  2. Jens Krause

    Present address: Chair of Genetics of Prokaryotes, Faculty of Biology, University of Bielefeld, 33501, Bielefeld, Germany

Authors and Affiliations

  1. Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, 48149, Münster, Germany

    Roland Klassen, Sabrina Wemhoff, Jens Krause & Friedhelm Meinhardt

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  1. Roland Klassen
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  2. Sabrina Wemhoff
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Correspondence to Friedhelm Meinhardt.

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Communicated by A. Aguilera.

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Klassen, R., Wemhoff, S., Krause, J. et al. DNA repair defects sensitize cells to anticodon nuclease yeast killer toxins. Mol Genet Genomics 285, 185–195 (2011). https://doi.org/10.1007/s00438-010-0597-5

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  • DOI: https://doi.org/10.1007/s00438-010-0597-5

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