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Vacuolar proteins in development of yeast colonies
Trubitsyna, Yana ; Palková, Zdena (advisor) ; Heidingsfeld, Olga (referee)
The laboratory strains of yeast Saccharomyces Cerevisiae form colonies which can differentiate into two main cell subpopulations. U and L cells demonstrate different morphology, metabolism and stress-resistance. It was also proved that some of metabolic pathways in U cells are a similar to ones in tumor cells. The unique metabolism is activated in U cells; the TORC1 is active in these cells together with autophagy and glycogen accumulation, which are characteristic for cells with inactivated TORC1. CORVET and HOPS complexes together with vacuolar ATPase are involved in processes related to vacuolar fusion and trafficking. Also, these complexes contribute to the regulation of TORC1 activity. Vam6p is a subunit of HOPS complex and it is also involved in regulation of TORC1 acting as GEF for Gtr1p GTPase, which activates TORC1. The aim of this study was to outline whether selected subunits of mentioned complexes affect TORC1 activity in U cells. Further aim was to confirm the effect of Vam6p on selected proteins production. These proteins were chosen based on results of proteomic analysis performed in our laboratory. In order to investigate possible effects of proteins of interest absence on colonies' morphology, strains deleted in selected genes were prepared (VPS3, VPS8, VPS33, VPS41, VPH2, VAC7 a...
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Vacuolar proteins in development of yeast colonies
Trubitsyna, Yana ; Palková, Zdena (advisor) ; Heidingsfeld, Olga (referee)
The laboratory strains of yeast Saccharomyces Cerevisiae form colonies which can differentiate into two main cell subpopulations. U and L cells demonstrate different morphology, metabolism and stress-resistance. It was also proved that some of metabolic pathways in U cells are a similar to ones in tumor cells. The unique metabolism is activated in U cells; the TORC1 is active in these cells together with autophagy and glycogen accumulation, which are characteristic for cells with inactivated TORC1. CORVET and HOPS complexes together with vacuolar ATPase are involved in processes related to vacuolar fusion and trafficking. Also, these complexes contribute to the regulation of TORC1 activity. Vam6p is a subunit of HOPS complex and it is also involved in regulation of TORC1 acting as GEF for Gtr1p GTPase, which activates TORC1. The aim of this study was to outline whether selected subunits of mentioned complexes affect TORC1 activity in U cells. Further aim was to confirm the effect of Vam6p on selected proteins production. These proteins were chosen based on results of proteomic analysis performed in our laboratory. In order to investigate possible effects of proteins of interest absence on colonies' morphology, strains deleted in selected genes were prepared (VPS3, VPS8, VPS33, VPS41, VPH2, VAC7 a...
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