title: Novel yeast killer toxins provoke S-phase arrest and DNA damage checkpoint activation.
abstract: Certain strains of Pichia acaciae and Wingea robertsiae (synonym Debaryomyces robertsiae) harbour extranuclear genetic elements that confer a killer phenotype to their host. Such killer plasmids (pPac1-2 of P. acaciae and pWR1A of W. robertsiae) were sequenced and compared with the zymocin encoding pGKL1 of Kluyveromyces lactis. Both new elements were found to be closely related to each other, but they are only partly similar to pGKL1. As for the latter, they encode functions mediating binding of the toxin to the target cell's chitin and a hydrophobic region potentially involved in uptake of a toxin subunit by target cells. Consistently, mutations affecting the target cell's major chitin synthase (Chs3) protect it from toxin action. Heterologous intracellular expression of respective open reading frames identified cell cycle-arresting toxin subunits deviating structurally from the likewise imported gamma-subunit of the K. lactis zymocin. Accordingly, toxicity of both P. acaciae and Wingea toxins was shown to be independent of RNA polymerase II Elongator, which is indispensable for zymocin action. Thus, P. acaciae and Wingea toxins differ in their mode of action from the G1-arresting zymocin. Fluorescence-activated cell sorting analysis and determination of budding indices have proved that such novel toxins mediate cell cycle arrest post-G1 during the S phase. Concomitantly, the DNA damage checkpoint kinase Rad53 is phosphorylated. As a mutant carrying the checkpoint-deficient allele rad53-11 displays toxin hypersensitivity, damage checkpoint activation apparently contributes to coping with toxin stress, rather than being functionally implemented in toxin action.
authors.affiliation: Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, Corrensstr. 3, D-48149 Münster, Germany.
authors.forename: Roland
authors.initials: R
authors.lastname: Klassen
authors.forename: Sabine
authors.initials: S
authors.lastname: Teichert
authors.forename: Friedhelm
authors.initials: F
authors.lastname: Meinhardt
mesh_terms: D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects*; D055646:Killer Factors, Yeast; D007716:Kluyveromyces / Q000737:chemistry; D009183:Mycotoxins / Q000737:chemistry / Q000494:pharmacology*; D010843:Pichia / Q000737:chemistry; D016196:S Phase / Q000187:drug effects*; D015003:Yeasts / Q000166:cytology / Q000187:drug effects*
chemical_list: D055646:Killer Factors, Yeast; D009183:Mycotoxins; C058975:zymocin; C087906:killer toxin, Pichia