Immunity factors for two related tRNAGln targeting killer toxins distinguish cognate and non-cognate toxic subunits.
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 tRNA(Gln) increases DrT resistance and the amount of tRNA(Gln) 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 tRNA(Gln) but not tRNA(Glu) 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.
Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, Corrensstr. 3, 48149, Münster, Germany, roland.klassen@uni-kassel.de.
Roland
R
Klassen
Alene
A
Kast
Guido
G
Wünsche
John P
JP
Paluszynski
Sabrina
S
Wemhoff
Friedhelm
F
Meinhardt
D004722:Endoribonucleases / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D023061:Gene Order; D007109:Immunity / Q000235:genetics; D007155:Immunologic Factors / Q000235:genetics* / Q000378:metabolism; D055646:Killer Factors, Yeast / Q000235:genetics* / Q000378:metabolism*; D059367:RNA Cleavage; D012351:RNA, Transfer, Gln / Q000235:genetics* / Q000378:metabolism*
D005656:Fungal Proteins; D007155:Immunologic Factors; D055646:Killer Factors, Yeast; D012351:RNA, Transfer, Gln; D004722:Endoribonucleases