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Immunity factors for two related tRNAGln targeting killer toxins distinguish cognate and non-cognate toxic subunits

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Abstract

The cytoplasmic virus-like element pWR1A from Debaryomyces robertsiae encodes a toxin (DrT) with similarities to the Pichia acaciae killer toxin PaT, which acts by importing a toxin subunit (PaOrf2) with tRNA anticodon nuclease activity into target cells. As for PaT, loss of the tRNA methyltransferase Trm9 or overexpression of tRNAGln increases DrT resistance and the amount of tRNAGln is reduced upon toxin exposure or upon induced intracellular expression of the toxic DrT subunit gene DrORF3, indicating DrT and PaT to share the same in vivo target. Consistent with a specific tRNase activity of DrOrf3, the protein cleaves tRNAGln but not tRNAGlu in vitro. Heterologous cytoplasmic expression identified DrOrf5 as the DrT specific immunity factor; it confers resistance to exogenous DrT as well as to intracellular expression of DrOrf3 and prevents tRNA depletion by the latter. The PaT immunity factor PaOrf4, a homologue of DrOrf5 disables intracellular action of both toxins. However, the DrT protection level mediated by PaOrf4 is reduced compared to DrOrf5, implying a recognition mechanism for the cognate toxic subunit, leading to incomplete toxicity suppression of similar, but non-cognate toxic subunits.

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Author notes

  1. John P. Paluszynski

    Present address: Division of Infectious Diseases, Cincinnati Children’s Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, OH, 45229, USA

Authors and Affiliations

  1. Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, Corrensstr. 3, 48149, Münster, Germany

    Roland Klassen, Alene Kast, Guido Wünsche, John P. Paluszynski, Sabrina Wemhoff & Friedhelm Meinhardt

  2. Fachgebiet Mikrobiologie, Universität Kassel, Heinrich-Plett-Str. 40, 34132, Kassel, Germany

    Roland Klassen

Authors
  1. Roland Klassen
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  2. Alene Kast
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  3. Guido Wünsche
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  4. John P. Paluszynski
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  5. Sabrina Wemhoff
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  6. Friedhelm Meinhardt
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Correspondence to Roland Klassen or Friedhelm Meinhardt.

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Communicated by S. Hohmann.

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Klassen, R., Kast, A., Wünsche, G. et al. Immunity factors for two related tRNAGln targeting killer toxins distinguish cognate and non-cognate toxic subunits. Curr Genet 60, 213–222 (2014). https://doi.org/10.1007/s00294-014-0426-1

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  • DOI: https://doi.org/10.1007/s00294-014-0426-1

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