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Antagonistic regulation by global transcription factors Tup1p, and Cyc8p of Flo11 and Flo11 -dependent phenotypes in wild yeast
Nguyen Van, Phu ; Palková, Zdena (advisor) ; Heidingsfeld, Olga (referee) ; Malcová, Ivana (referee)
Biofilms are a common mode of yeast growth in which cells adhere to each other and adhere to abiotic surfaces to form complex multicellular structures. Living together in biofilms provides cells with several beneficial features compared to planktonic cells. Undoubtedly, protection and resistance are advantages of life inside colony biofilms. Biofilms are found in many environments and play many important roles in commercial industries. However, biofilms can also be extremely dangerous in clinical settings. There is thus great interest in studying biofilms and how to eliminate them. In this study, we used wild yeast Saccharomyces cerevisiae colony biofilm as an ideal system to investigate potential functions of the yeast Cyc8-Tup1 transcriptional corepressor complex in the regulation of yeast adhesion, and biofilm formation on agar and at solid-liquid interfaces. Unexpectedly, we have found that Cyc8p and Tup1p antagonistically control the formation of structured biofilm colonies on agar and FLO11 expression. Cyc8p itself acts as a key repressor of FLO11, whereas Tup1p promotes the formation of biofilm colonies and induces FLO11 expression by inhibiting the repressive function of Cyc8p and preventing Flo11p degradation possibly by inhibiting an extracellular protease. In addition, other features...
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Cell polarity establishment and changes during Saccharomyces cerevisiae cell cycle
Luxová, Pavla ; Malcová, Ivana (advisor) ; Šťovíček, Vratislav (referee)
Cell polarity can be defined as an asymmetric organization and distribution of biomolecules, cellular organelles and structures which are important for many cellular processes. Cell polarity establishment is essential for the proper development of all organisms. This work focuses on main mechanisms of cell polarity establishment, its maintenance and changes during Saccharomyces cerevisiae cell cycle. Budding yeast is one of the preferred model organism. Bud site selection is determined by the spatial landmarks which are accumulated at the previous division site. The spatial landmarks are recognized by Rho GTPases which act on their effectors and thus affect the actin cytoskeleton and septins. These structures are essential for polarized bud growth that is coordinated with the cell cycle. Newborn cells arising after the bud separation from the mother cell at the end of each cycle are able to undergo many more division cycles than their mothers what is a new challenge to study cell polarity in terms of cell aging.
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Molecular base of plant HSP90-MT interaction
Benáková, Martina ; Krtková, Jana (advisor) ; Malcová, Ivana (referee)
Microtubules (MTs) are one of the essential cell structure that participate in a number of key events in the plant cells and their properties and functions are influenced and modified by many other proteins. These proteins belong to a group of microtubule- associated proteins (MAPs, microtubule-associated proteins). One of the MAPs, the molecular chaperone Hsp90, examines and fulfills a large number of different functions in the cell. Its colocalization with MTs has been demonstrated previously by Freudenreich and Nick (1998) and Petrášek et al. (1998). However, direct interaction with MTs was described only recently using cosedimentation assay. The specific cytosolic isoform of tobacco Hsp90 bound to MTs was called Hsp90_MT due to its ability to bind MTs. It has been also found that the binding to MTs is independent on the activity of ATP (Krtková et al., 2012). The authors also described a positive effect of Hsp90_MT on MT recovery after their exposure to cold stress. Although MT cytoskeleton dynamics is influenced by a large number of MAPs, it is surprising that the molecular mechanism of MAPs interaction with MTs and their MT-binding domains have not been described yet. Therefore, we decided to determine the tobacco Hsp90_MT MT-binding domain by production of a set of recombinant proteins...
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Analysis of Rpg1/eIF3a mutation in Saccharomyces cerevisiae.
Luxová, Pavla ; Malcová, Ivana (advisor) ; Binarová, Pavla (referee)
Rpg1/Tif32/eIF3a is an essential and the largest subunit of translation initiation factor eIF3 in yeast Saccharomyces cerevisiae. Besides interactions within the eIF3 complex it has been shown to interact with microtubules. Preliminary data of the laboratory obtained using strains of the W303 genetic background indicated that there is a synthetic phenotype between rpg1-2 mutant and microtubule inhibitor nocodazole. Aim of this work to elucidate this "microtubule phenotype" of the rpg1-2 mutant and its dependency on used genetic background. I confirmed that independently on genetic background (W303, BY, SEY) all mutants rpg1-1, rpg1-2 and rpg1-3 were temperature- sensitive. I found that in contrast to published data on rpg1 mutants of the W303 background these mutants of the BY and the SEY backgrounds do not arrest the cell cycle in G1 phase during cultivation at the restrictive temperature (37řC, 4 hours). In addition, all three mutants did not show an increased sensitivity to benomyl and none of them affects microtubule rearrangement after a release of cells from the nocodazole treatment. I constructed new strains with a combination of the BUB1 gene deletion with the particular rpg1 mutation. Phenotypic analyses of new double mutants revealed that simultaneous dis-function of Bub1 and Rpg1 results...
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The effect of antimicrobial peptides on Saccharomyces cerevisiae and other yeast species
Makarova, Anna-Marie ; Hodek, Petr (advisor) ; Malcová, Ivana (referee)
The increased use of antibiotics, antifungal agents and disinfectants in the last decades has resulted in development of microbial resistance to these drugs. Candida species are the fourth most common cause of hospital-acquired bloodstream infection and kill 40% of those patients. Natural antimicrobial peptides are promising candidates for the development of new agents to treat yeast and bacterial infections, as their presumed mechanism of action differs significantly from the mechanism of action of current drugs. This work is focused on several peptides isolated from the venom of wild bees and their synthetic analogues and the identification of the most effective ones against non-pathogenic Saccharomyces cerevisiae and several pathogenic Candida species. Antifungal activity of eight cationic antimicrobial peptides was tested and compared under various conditions. The overall susceptibility of pathogenic yeast species to currently used antifungal drugs and the antimicrobial peptides was screened with the aim to identify potential synergistic and species-specific effects. The effect of antimicrobial peptides on membrane potential was measured by a fluorescent probe (diS-C3(3)), and the relative hyperpolarization of plasma membrane was shown for each peptide. The effect of antimicrobial peptides on...
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Antagonistic regulation by global transcription factors Tup1p, and Cyc8p of Flo11 and Flo11 -dependent phenotypes in wild yeast
Nguyen Van, Phu ; Palková, Zdena (advisor) ; Heidingsfeld, Olga (referee) ; Malcová, Ivana (referee) ; Demnerová, Kateřina (referee)
Biofilms are a common mode of yeast growth in which cells adhere to each other and adhere to biotic and abiotic surfaces to form complex multicellular structures. Living together in biofilms provides cells with several benefits, compared to planktonic cells such as protection and resistance to antimicrobials, environmental stresses and host immune attacks. Biofilms may play many important roles in commercial industries. But they are considered to be extremely dangerous in clinical settings. There is thus great interest in studying biofilms and how to eliminate them. In this study, we used wild yeast Saccharomyces cerevisiae colony biofilm as an ideal system to investigate potential functions of the yeast Cyc8p-Tup1p transcriptional corepressor complex in the regulation of yeast adhesion and biofilm formation on agar and at solid-liquid interfaces. Unexpectedly, we found that Cyc8p and Tup1p antagonistically control FLO11 expression and the formation of structured biofilm colonies on agar. Cyc8p itself acts as a key repressor of FLO11 and biofilm colony formation, whereas Tup1p promotes the formation of biofilm colonies and induces FLO11 expression by inhibiting the repressive function of Cyc8p and preventing Flo11p degradation possibly by inhibiting an extracellular protease. Other typical features...
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Antagonistic regulation by global transcription factors Tup1p, and Cyc8p of Flo11 and Flo11 -dependent phenotypes in wild yeast
Nguyen Van, Phu ; Palková, Zdena (advisor) ; Heidingsfeld, Olga (referee) ; Malcová, Ivana (referee)
Biofilms are a common mode of yeast growth in which cells adhere to each other and adhere to abiotic surfaces to form complex multicellular structures. Living together in biofilms provides cells with several beneficial features compared to planktonic cells. Undoubtedly, protection and resistance are advantages of life inside colony biofilms. Biofilms are found in many environments and play many important roles in commercial industries. However, biofilms can also be extremely dangerous in clinical settings. There is thus great interest in studying biofilms and how to eliminate them. In this study, we used wild yeast Saccharomyces cerevisiae colony biofilm as an ideal system to investigate potential functions of the yeast Cyc8-Tup1 transcriptional corepressor complex in the regulation of yeast adhesion, and biofilm formation on agar and at solid-liquid interfaces. Unexpectedly, we have found that Cyc8p and Tup1p antagonistically control the formation of structured biofilm colonies on agar and FLO11 expression. Cyc8p itself acts as a key repressor of FLO11, whereas Tup1p promotes the formation of biofilm colonies and induces FLO11 expression by inhibiting the repressive function of Cyc8p and preventing Flo11p degradation possibly by inhibiting an extracellular protease. In addition, other features...
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Regulation of yeast chronological ageing
Fedorová, Viktória ; Palková, Zdena (advisor) ; Malcová, Ivana (referee)
After exhausting the nutrients from medium, yeast cells enter the stacionary phase, in which they adapt their metabolism and physiological functions and undergo chronological ageing characterized by a non-dividing, or only slowly-dividing cell state. This type of ageing is comparable with post-mitotic ageing of cells in higher eukaryotes. Chronologically aged yeast populations form two subpopulations of interdependent cells in liquid cultures as well as in colonies grown on solid media. Chronological ageing is understood as a period of time during which yeast population is able to survive in the stacionary phase, and is regulated by caloric restriction and evolutionary conserved pathways, which respond to various substances, such as carbon or amino acids. Metabolites produced by cellular processes or various chemicals found in the natural environment of these single-celled organisms also have regulatory effects on yeast chronological ageing. Key words: yeast, Saccharomyces cerevisiae, stationary phase, chronological ageing of yeast, cell differentiation, regulatory pathways, metabolites
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The effect of antimicrobial peptides on Saccharomyces cerevisiae and other yeast species
Makarova, Anna-Marie ; Hodek, Petr (advisor) ; Malcová, Ivana (referee)
The increased use of antibiotics, antifungal agents and disinfectants in the last decades has resulted in development of microbial resistance to these drugs. Candida species are the fourth most common cause of hospital-acquired bloodstream infection and kill 40% of those patients. Natural antimicrobial peptides are promising candidates for the development of new agents to treat yeast and bacterial infections, as their presumed mechanism of action differs significantly from the mechanism of action of current drugs. This work is focused on several peptides isolated from the venom of wild bees and their synthetic analogues and the identification of the most effective ones against non-pathogenic Saccharomyces cerevisiae and several pathogenic Candida species. Antifungal activity of eight cationic antimicrobial peptides was tested and compared under various conditions. The overall susceptibility of pathogenic yeast species to currently used antifungal drugs and the antimicrobial peptides was screened with the aim to identify potential synergistic and species-specific effects. The effect of antimicrobial peptides on membrane potential was measured by a fluorescent probe (diS-C3(3)), and the relative hyperpolarization of plasma membrane was shown for each peptide. The effect of antimicrobial peptides on...
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Biochemical and molecular basis of specific properties of non-conventional osmotolerant yeast
Dušková, Michala ; Sychrová, Hana (advisor) ; Malcová, Ivana (referee) ; Janderová, Blanka (referee)
Unicellular organisms such as yeasts are permanently exposed to environmental changes, especially to changes of the concentration of osmotically active substances. Yeast species, which are able to cope with these changes properly, are called osmotolerant (Zygosaccharomyces rouxii, Pichia sorbitophila, Debaryomyces hansenii, etc.). The osmotolerance of yeasts depends on many physiological parameters but the most important is the efficient metabolism of internal osmolytes. This role is played by the small molecule of glycerol in most yeast species. Yeasts have to release the surplus of glycerol during the hypoosmotic shock. On the other hand, the hyperosmotic conditions require synthesis and accumulation of this small compound in a high quantity. The model yeast Saccharomyces cerevisiae has two systems transporting glycerol. First of them is the ScFps1 channel which is important mainly for releasing of glycerol during hypoosmotic shock. The second transporter, called ScStl1, is employed in accumulation of glycerol in case of hyperosmotic conditions. Detailed study of physiological properties of the three above mentioned species was performed within this work. Although they do not differ in basic morphological parameters from S. cerevisiae, their difference lies in the ability to survive desiccation,...
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