title: Architecture of developing multicellular yeast colony: spatio-temporal expression of Ato1p ammonium exporter.
abstract: Yeasts, when growing on solid surfaces, form organized multicellular structures, colonies, in which cells differentiate and thus possess different functions and undergo dissimilar fate. Understanding the principles involved in the formation of these structures requires new approaches that allow the study of individual cells directly in situ without needing to remove them from the microbial community. Here we introduced a new approach to the analysis of whole yeast microcolonies either containing specific proteins labelled by fluorescent proteins or stained with specific dyes, by two-photon excitation confocal microscopy. It revealed that the colonies are covered with a thin protective skin-like surface cell layer which blocks penetration of harmful compounds. The cells forming the layer are tightly connected via cell walls, the presence of which is essential for keeping of protective layer function. Viewing the colonies from different angles allowed us to reconstruct a three-dimensional profile of the cells producing ammonium exporter Ato1p within developing microcolonies growing either as individuals or within a group of microcolonies. We show that neighbouring microcolonies coordinate production of Ato1p-GFP. Ato1p itself appears synchronously in cells, which do not originate from the same ancestor, but occupy specific position within the colony.
authors.affiliation: Institute of Microbiology AS CR, v.v.i., Vídenská 1083, 142 20 Prague 4, Czech Republic.
authors.forename: Libuse
authors.initials: L
authors.lastname: Váchová
authors.forename: Oleksandr
authors.initials: O
authors.lastname: Chernyavskiy
authors.forename: Dita
authors.initials: D
authors.lastname: Strachotová
authors.forename: Paolo
authors.initials: P
authors.lastname: Bianchini
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Burdíková
authors.forename: Ivana
authors.initials: I
authors.lastname: Fercíková
authors.forename: Lucie
authors.initials: L
authors.lastname: Kubínová
authors.forename: Zdena
authors.initials: Z
authors.lastname: Palková
mesh_terms: D015966:Gene Expression Regulation, Fungal; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000096:biosynthesis*; D018613:Microscopy, Confocal / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000254:growth & development*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis*; D013194:Staining and Labeling / Q000379:methods; D013997:Time Factors
chemical_list: C520069:ATO1 protein, S cerevisiae; D026901:Membrane Transport Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins