title: FLO11 Gene Is Involved in the Interaction of Flor Strains of Saccharomyces cerevisiae with a Biofilm-Promoting Synthetic Hexapeptide.
abstract: Saccharomyces cerevisiae "flor" yeasts have the ability to form a buoyant biofilm at the air-liquid interface of wine. The formation of biofilm, also called velum, depends on FLO11 gene length and expression. FLO11 encodes a cell wall mucin-like glycoprotein with a highly O-glycosylated central domain and an N-terminal domain that mediates homotypic adhesion between cells. In the present study, we tested previously known antimicrobial peptides with different mechanisms of antimicrobial action for their effect on the viability and ability to form biofilm of S. cerevisiae flor strains. We found that PAF26, a synthetic tryptophan-rich cationic hexapeptide that belongs to the class of antimicrobial peptides with cell-penetrating properties, but not other antimicrobial peptides, enhanced biofilm formation without affecting cell viability in ethanol-rich medium. The PAF26 biofilm enhancement required a functional FLO11 but was not accompanied by increased FLO11 expression. Moreover, fluorescence microscopy and flow cytometry analyses showed that the PAF26 peptide binds flor yeast cells and that a flo11 gene knockout mutant lost the ability to bind PAF26 but not P113, a different cell-penetrating antifungal peptide, demonstrating that the FLO11 gene is selectively involved in the interaction of PAF26 with cells. Taken together, our data suggest that the cationic and hydrophobic PAF26 hexapeptide interacts with the hydrophobic and negatively charged cell wall, favoring Flo11p-mediated cell-to-cell adhesion and thus increasing biofilm biomass formation. The results are consistent with previous data that point to glycosylated mucin-like proteins at the fungal cell wall as potential interacting partners for antifungal peptides.
authors.affiliation: Dipartimento di Agraria, University of Sassari, Sassari, Italy.
authors.forename: Marc
authors.initials: M
authors.lastname: Bou Zeidan
authors.forename: Lourdes
authors.initials: L
authors.lastname: Carmona
authors.forename: Severino
authors.initials: S
authors.lastname: Zara
authors.forename: Jose F
authors.initials: JF
authors.lastname: Marcos
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000378:metabolism*; D018441:Biofilms / Q000254:growth & development*; D003470:Culture Media / Q000737:chemistry; D005434:Flow Cytometry; D055786:Gene Knockout Techniques; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D050296:Microbial Viability; D008856:Microscopy, Fluorescence; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D023181:Antimicrobial Cationic Peptides; D003470:Culture Media; C110690:FLO11 protein, S cerevisiae; D008562:Membrane Glycoproteins; D029701:Saccharomyces cerevisiae Proteins
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