|
|
Long term sublethal stress in colonies of Saccharomyces cerevisiae deleted in WHI3
Krampotová, Ester ; Schierová, Michaela (advisor) ; Dušková, Michala (referee)
This diploma thesis is devoted to the topic of the development of natural non-pathogenic yeast colonies in non-laboratory diploid strain of Saccharomyces cerevisae with deletion WHI3, during long-term mild stress. The absence of Whi3 causes significantly higher senzitivity to medium composition relative to the parental strain, thus whi3Δ /whi3Δ colonies are a good model for study of the virulence induction due to environmental stress in pathogenic yeasts. Deletion of WHI3 in the BR-F strain results in a significant suppression of the ability to form structured colonies caused mainly by the reduced level of Flo11. The absence of Whi3 also has a negative effect on the expression of other genes involved in the stress response. The aim of the work is to determine whether changes in the expression of genes encoding stress proteins induced by sublethal doses of the inhibitor are dependent on Whi3, Yap6 and Mpt5 proteins. To induce stress in yeast, we used NaCl or CdSO4. The YAP6 and MPT5 genes encode regulatory proteins involved in the stress response, which expression is under control of Whi3. In whi3Δ /whi3Δ , the level of Hsp26, Tsa1, Pab1, and Gre2 was lower than in the parental strain. The Yap6 protein affects the response to the presence of Cd2+ in the medium, although according to SGD, only its...
|
|
|
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...
|
|
|
Factors influencing cell adhesion and the formation of yeast biofilms on solid surfaces
Světlíková, Daniela ; Palková, Zdena (advisor) ; Groušl, Tomáš (referee)
Yeasts are unicellular microorganisms that naturally aggregate to form multicellular structures. One of these formations is biofilm. The key properties of biofilm in S. cerevisiae are determined by the expression of the FLO11 gene. The large FLO11 gene promoter is the target of several pathways and factors, resulting in the complex regulation of this gene. Among the regulators of the FLO11 gene are the conserved Cyc8p and Tup1p proteins, which play a role in the regulation of many processes, especially as a corepressor complex. In previous years, our laboratory has demonstrated an antagonistic effect of the Cyc8p a Tup1p regulators on adhesion of BRF strain (Nguyen et al., 2018). In my work, I investigated whether this regulation is also valid in the selected clinical isolate S. cerevisiae YJM320 and whether, as in the BRF strain, positive regulation of FLO11 via Tup1 and negative regulation via Cyc8 exist here or whether their effect is different in this strain. The broader goal of my work was to identify the role of various factors on adhesion as the first step in biofilm formation. My work began by testing the effect of nutrients and temperature on the adhesion of strains BY, BRS and BRF and continued by testing the adhesion of selected clinical isolates of Saccharomyces cerevisiae and Candida...
|
|
|
Differentiation of yeast colonies: The role of selected transcription factors and metabolic proteins
Plocek, Vítězslav
5 Abstract Although yeasts are unicellular microorganisms, they form complex multicellular formations such as biofilms and colonies under natural conditions. Within these structures, processes such as cell differentiation, specialization by particular cell populations and cell signalling, which are typical of multicellular organisms, take place. The literature introduction to this thesis summarizes current knowledge regarding the development of biofilms and colonies, in particular those of the model organism, Saccharomyces cerevisiae, and some selected regulations that are important for the formation of multicellular structures. In the results section, I focus on two lines of research. The first is directed towards mechanisms, involved in the formation of multicellular structures. In studying the formation of SLI biofilms (biofilms at the solid/liquid interface), we have documented the antagonistic role of the regulators CYC8 and TUP1 in their formation and have also described the effect of the presence of glucose on the development and stability of SLI biofilms of strain BR-F. During this study we[D1] have developed an imaging method that allows us to prepare and observe the internal structure (vertical cross-section) of SLI biofilms, as well as the growth of unattached cells, under physiological...
|
|
|
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...
|
|
|
Differentiation of yeast colonies: The role of selected transcription factors and metabolic proteins
Plocek, Vítězslav ; Palková, Zdena (advisor) ; Heidingsfeld, Olga (referee) ; Sychrová, Hana (referee)
5 Abstract Although yeasts are unicellular microorganisms, they form complex multicellular formations such as biofilms and colonies under natural conditions. Within these structures, processes such as cell differentiation, specialization by particular cell populations and cell signalling, which are typical of multicellular organisms, take place. The literature introduction to this thesis summarizes current knowledge regarding the development of biofilms and colonies, in particular those of the model organism, Saccharomyces cerevisiae, and some selected regulations that are important for the formation of multicellular structures. In the results section, I focus on two lines of research. The first is directed towards mechanisms, involved in the formation of multicellular structures. In studying the formation of SLI biofilms (biofilms at the solid/liquid interface), we have documented the antagonistic role of the regulators CYC8 and TUP1 in their formation and have also described the effect of the presence of glucose on the development and stability of SLI biofilms of strain BR-F. During this study we[D1] have developed an imaging method that allows us to prepare and observe the internal structure (vertical cross-section) of SLI biofilms, as well as the growth of unattached cells, under physiological...
|
|
|
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...
|
|
|
Differentiation of yeast colonies and development of new approaches to monitor oxygen and nutrient availability
Vopálenská, Irena ; Janderová, Blanka (advisor) ; Demnerová, Kateřina (referee) ; Pichová, Iva (referee)
Yeast Saccharomyces cerevisiae as an unicellular organism is one of the best-studied experimental organisms. It is an important model organism for the study of intracellular processes of eukaryotic cells. Yeasts are also social organisms with cell-to-cell communication able to form organized multicellular structures (colonies and biofilms). Yeast and other microorganisms in nature prefer to form colonies on solid substrates rather than to grow as "planktonic" single cells (Palková, 2004; Wimpenny, 2009). The yeast S. cerevisiae typically forms colonies, biofilms were described only rarely. Yeast colonies exhibit an organized morphological pattern characteristic of each particular yeast strain (Kocková-Kratochvílová, 1982). This work is focusing on morphology and differentiation of the S. cerevisiae colonies of common laboratory strains forming less structured colonies, and strains of the Σ1278b genetic background forming highly structured "fluffy" colonies. It shows that polarized budding pattern and especially cell ability to form aggregates enable development of structured morphology. During development of "fluffy" colonies two differently regulated events of dimorphic switch from yeast form to filamentous growth occur. One of these events is dependent on the surface glycoprotein, Flo11p flocculin. This...
|
guest ::
