title: Yeast β-1,6-glucan is a primary target for the Saccharomyces cerevisiae K2 toxin.
abstract: Certain Saccharomyces cerevisiae strains secrete different killer proteins of double-stranded-RNA origin. These proteins confer a growth advantage to their host by increasing its survival. K2 toxin affects the target cell by binding to the cell surface, disrupting the plasma membrane integrity, and inducing ion leakage. In this study, we determined that K2 toxin saturates the yeast cell surface receptors in 10 min. The apparent amount of K2 toxin, bound to a single cell of wild type yeast under saturating conditions, was estimated to be 435 to 460 molecules. It was found that an increased level of β-1,6-glucan directly correlates with the number of toxin molecules bound, thereby impacting the morphology and determining the fate of the yeast cell. We observed that the binding of K2 toxin to the yeast surface receptors proceeds in a similar manner as in case of the related K1 killer protein. It was demonstrated that the externally supplied pustulan, a poly-β-1,6-glucan, but not the glucans bearing other linkage types (such as laminarin, chitin, and pullulan) efficiently inhibits the K2 toxin killing activity. In addition, the analysis of toxin binding to the intact cells and spheroplasts confirmed that majority of K2 protein molecules attach to the β-1,6-glucan, rather than the plasma membrane-localized receptors. Taken together, our results reveal that β-1,6-glucan is a primary target of K2 toxin and is important for the execution of its killing property.
authors.affiliation: Laboratory of Genetics, Institute of Botany, Nature Research Centre, Vilnius, Lithuania.
authors.forename: Juliana
authors.initials: J
authors.lastname: Lukša
authors.affiliation: Laboratory of Genetics, Institute of Botany, Nature Research Centre, Vilnius, Lithuania.
authors.forename: Monika
authors.initials: M
authors.lastname: Podoliankaitė
authors.affiliation: Laboratory of Genetics, Institute of Botany, Nature Research Centre, Vilnius, Lithuania.
authors.forename: Iglė
authors.initials: I
authors.lastname: Vepštaitė
authors.affiliation: Laboratory of Genetics, Institute of Botany, Nature Research Centre, Vilnius, Lithuania.
authors.forename: Živilė
authors.initials: Ž
authors.lastname: Strazdaitė-Žielienė
authors.affiliation: Laboratory of Genetics, Institute of Botany, Nature Research Centre, Vilnius, Lithuania Department of Molecular Microbiology and Biotechnology, Institute of Biochemistry, Vilnius University, Vilnius, Lithuania.
authors.forename: Jaunius
authors.initials: J
authors.lastname: Urbonavičius
authors.affiliation: Laboratory of Genetics, Institute of Botany, Nature Research Centre, Vilnius, Lithuania elena.serviene@botanika.lt.
authors.forename: Elena
authors.initials: E
authors.lastname: Servienė
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism; D002686:Chitin / Q000378:metabolism; D005936:Glucans / Q000378:metabolism; D055646:Killer Factors, Yeast / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D013104:Spheroplasts; D047071:beta-Glucans / Q000378:metabolism*
chemical_list: D005936:Glucans; C073585:K2 killer toxin; D055646:Killer Factors, Yeast; D011134:Polysaccharides; D029701:Saccharomyces cerevisiae Proteins; D047071:beta-Glucans; D002686:Chitin; C002076:pustulan; C064197:beta-1,6-glucan; C009109:pullulan; C008247:laminaran