PhD thesis will be part of the project focused on studies of molecular and cellular mechanisms involved in development of yeast colonies. Previous research revealed prominent differentiation of Saccharomyces cerevisiae colonies (Mol Cell 46: 436-48, 2012; Ox Med Cell Longev, doi:10.1155/2013/102485, 2013, Cell Cycle 14: 3488-3497, 2015). Chronologically aged S. cerevisiae colonies are composed of the two major cell-types: U cells in upper regions that activate adaptive metabolism and gain longevity phenotype and L cells in lower regions that behave as starved and stressed cells and seem to provide nutrients to U cells. Recent research revealed important role of signaling from mitochondria in development and longevity of the specifically localized colonial cell subpopulations as well as the presence of at least 3 different branches of mitochondrial signaling in differentiated colonies (Oncotarget 7: 15299-314, 2016). The subject of PhD thesis is based on these findings and will focus on the comparison of development and differentiation of S. cerevisiae colonies and colonies of yeast pathogens such as Candida glabrata. In particular, C. glabrata orthologues of genes that are specifically expressed in differentiated cell subpopulations in S. cerevisiae colonies will be identified and subsequently used to find and characterize similar subpopulations in C. glabrata colonies and biofilms. Emphasis will be given to analysis of mitochondrial signalling in colonies and biofilms of pathogenic strains of C. glabrata and S. cerevisiae. Spectrum of different approaches will be used during the project, including construction of knockout strains and strains with proteins labelled with fluorescent tags, in situ characterization of specific cell subpopulations using fluorescent and confocal microscopy and separation of subtle cell subpopulations of the colonies for analyses by different molecular and cellular biology techniques (including MS-MS proteomics and transcriptomics).
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