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CUG Codon in Pathogenic Yeasts of the Genus Candida
Marečková, Lucie ; Heidingsfeld, Olga (advisor) ; Půta, František (referee)
2. Abstract In many Candida species the standard leucine CUG codon is translated as a serine, although not in 100% cases. This dual specifity of the CUG codon has evolved through a mechanism that required codon ambiguity mediated by a unique tRNACAG, which is in vitro aminoacylated more often by serine than by leucine. This codon ambiguity has been tolerated for more than 170 million years. The explanation at least for now is that the CUG codon reassignment could have generated genetic diversity that facilitated occurrence of new phenotypes resistant to stress. Beside this, an important step was to reduce negative impact of the codon ambiguity by crucial mutations in the structure of the ser-tRNACAG. Candida species became a valuable experimental model for elucidation of the genetic code changes. While consequences of the CUG codon reassignment have been extensively studied in Candida albicans, this topic has not yet been addressed in Candida parapsilosis. Solving the structure of C. parapsilosis secreted proteinase Sapp1p provided a tool to carry out a "case study" of possible effects of the CUG codon ambiguity. The SAPP1 gene contains one CUG codon, and the respective serine is located on the loop in the close proximity of the active site of the proteinase.
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Detection of Sap2 in the secretome of Candida albicans cell wall and secretory mutants
Kollárová, Nikola ; Janďourek, Ondřej (advisor) ; Konečná, Klára (referee)
Candidate: Nikola Kollárová Title of diploma thesis: Detection of Sap2 in the secretome of Candida albicans cell wall and secretory mutants Charles University, Faculty of Pharmacy in Hradec Králové, Department of Biological and Medicinal Sciences Complutense University of Madrid, Faculty of Pharmacy, Department of Microbiology II Study program: Pharmacy Backgound: The aim of this diploma thesis was to search for C. albicans proteins involved in the secretion of the secreted aspartyl proteinase 2 enzyme (Sap2) evaluating the ability to degrade BSA (bovine serum albumin) as a source of nitrogen in several cell wall and secretory mutants of C. albicans. The work was carried out at the Department of Microbiology II, Faculty of Pharmacy, Complutense University of Madrid. Methods: The supernatant samples of several Candida albicans mutants were tested by SDS-PAGE electrophoresis and stained. Bands corresponding to BSA were observed and compared to controls. The other method was counted with 96-well plate. Results: The correlation between optical density and degradation of BSA was observed. Some mutants with disability to degrade BSA were found in a pilot screening of the ability to degrade BSA using 96-well plate method. That fact was confirmed by SDS-PAGE electrophoresis. C. albicans mutants showing...
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Analysis of protein cargo of extracellualr vesicles isolated from the yeast Candida albicans
Hlubučková, Lucie ; Konečná, Klára (advisor) ; Janďourek, Ondřej (referee)
Charles University Faculty of Pharmacy in Hradec Králové Study program: Pharmacy Candidate: Lucie Hlubučková Consultant: RNDr. Klára Konečná, Ph.D. Title of thesis: Analysis of protein cargo of extracellular vesicles isolated from the yeast Candida albicans Backgroung: The aim of this diploma thesis was to analyze the protein cargo carried in extracellular vesicles released from the yeast Candida albicans (C. albicans), which is one of the most important mycotic agens. Extracellular vesicles (EVs) are utilized as "transport vehicles", for the delivery of effector molecules into extracellular milieu. These molecules and primarilly proteins can play different roles in host-pathogen interactions. Proteins isolated from EVs and identified by proteomic approach were sorted into categories according to their molecular function and localization for the purpose of finding out, which proteins are predominantly distributed via extracellular vesicles into extracellular space after induction of nutrition starvation. Analysis of EVs protein cargo with focus on virulence factors could extend the knowledge about extracellular vesicles and their potential role in pathogenesis. Methods: Chosen C. albicans yeast strain was a clinical isolate strain isolated from a premenopausal women suffering from recurrent...
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Ammmonium transport in yeast
Faltýnková, Kateřina ; Palková, Zdena (advisor) ; Princová, Jarmila (referee)
Bachelor thesis - Kateřina Faltýnková - Ammmonium transport in yeast Ammonium and ammonia are an essential nutrient for every yeast cells, not only in metabolism, for example in amino acid synthesis, but also as signalling molecules that serve for communication between colonies or for the regulation of pseudohyphal growth. Transport of ammonia and ammonium ions requires active transport, which is provided by MEP permeases inside the cell likely by exporters ATO proteins out of the cell. In this work there are described families of genes MEP and ATO with main focus on their importance for uptake and export of ammonium ions by yeast and also the regulation of these two gene families in the yeast Saccharomyces cerevisiae, Candida albicans, Cryptococcus neoformans a Yarrowia lipolytica.
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Analysis of protein cargo of extracellualr vesicles isolated from the yeast Candida albicans
Hlubučková, Lucie ; Konečná, Klára (advisor) ; Janďourek, Ondřej (referee)
Charles University Faculty of Pharmacy in Hradec Králové Study program: Pharmacy Candidate: Lucie Hlubučková Consultant: RNDr. Klára Konečná, Ph.D. Title of thesis: Analysis of protein cargo of extracellular vesicles isolated from the yeast Candida albicans Backgroung: The aim of this diploma thesis was to analyze the protein cargo carried in extracellular vesicles released from the yeast Candida albicans (C. albicans), which is one of the most important mycotic agens. Extracellular vesicles (EVs) are utilized as "transport vehicles", for the delivery of effector molecules into extracellular milieu. These molecules and primarilly proteins can play different roles in host-pathogen interactions. Proteins isolated from EVs and identified by proteomic approach were sorted into categories according to their molecular function and localization for the purpose of finding out, which proteins are predominantly distributed via extracellular vesicles into extracellular space after induction of nutrition starvation. Analysis of EVs protein cargo with focus on virulence factors could extend the knowledge about extracellular vesicles and their potential role in pathogenesis. Methods: Chosen C. albicans yeast strain was a clinical isolate strain isolated from a premenopausal women suffering from recurrent...
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Detection of Sap2 in the secretome of Candida albicans cell wall and secretory mutants
Kollárová, Nikola ; Janďourek, Ondřej (advisor) ; Konečná, Klára (referee)
Candidate: Nikola Kollárová Title of diploma thesis: Detection of Sap2 in the secretome of Candida albicans cell wall and secretory mutants Charles University, Faculty of Pharmacy in Hradec Králové, Department of Biological and Medicinal Sciences Complutense University of Madrid, Faculty of Pharmacy, Department of Microbiology II Study program: Pharmacy Backgound: The aim of this diploma thesis was to search for C. albicans proteins involved in the secretion of the secreted aspartyl proteinase 2 enzyme (Sap2) evaluating the ability to degrade BSA (bovine serum albumin) as a source of nitrogen in several cell wall and secretory mutants of C. albicans. The work was carried out at the Department of Microbiology II, Faculty of Pharmacy, Complutense University of Madrid. Methods: The supernatant samples of several Candida albicans mutants were tested by SDS-PAGE electrophoresis and stained. Bands corresponding to BSA were observed and compared to controls. The other method was counted with 96-well plate. Results: The correlation between optical density and degradation of BSA was observed. Some mutants with disability to degrade BSA were found in a pilot screening of the ability to degrade BSA using 96-well plate method. That fact was confirmed by SDS-PAGE electrophoresis. C. albicans mutants showing...
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Detection of Sap2 in the secretome of Candida albicans cell wall and secretory mutants
Kollárová, Nikola ; Janďourek, Ondřej (advisor) ; Konečná, Klára (referee)
Candidate: Nikola Kollárová Title of diploma thesis: Detection of Sap2 in the secretome of Candida albicans cell wall and secretory mutants Charles University, Faculty of Pharmacy in Hradec Králové, Department of Biological and Medicinal Sciences Complutense University of Madrid, Faculty of Pharmacy, Department of Microbiology II Study program: Pharmacy Backgound: The aim of this diploma thesis was to search for C. albicans proteins involved in the secretion of the secreted aspartyl proteinase 2 enzyme (Sap2) evaluating the ability to degrade BSA (bovine serum albumin) as a source of nitrogen in several cell wall and secretory mutants of C. albicans. The work was carried out at the Department of Microbiology II, Faculty of Pharmacy, Complutense University of Madrid. Methods: The supernatant samples of several Candida albicans mutants were tested by SDS-PAGE electrophoresis and stained. Bands corresponding to BSA were observed and compared to controls. The other method was counted with 96-well plate. Results: The correlation between optical density and degradation of BSA was observed. Some mutants with disability to degrade BSA were found in a pilot screening of the ability to degrade BSA using 96-well plate method. That fact was confirmed by SDS-PAGE electrophoresis. C. albicans mutants showing...
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Tyrosol effects on minimal inhibition concentration of ATM in Candida albicans
Vaisová, Marcela ; Křivčíková, Lucie (advisor) ; Blažíčková, Kateřina (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Author: Marcela Vaisová Supervisor: Lucie Křivčíková Title of diploma thesis: Effect of tyrosol on minimal inhibitory concentration of antimycotics Previous studies concerning quorum sensing in Candida albicans discovered two quorum sensing molecules, farnesol and tyrosol. Tyrosol has been formerly confirmed as a substance which shortens the lag phase of Candida albicans cell cycle and stimulates its conversion to the hyphal form, farnesol stimulates blastospores. This work covers the effect of tyrosol combined with two antimycotics, fungistatic fluconazole and fungicidal amphotericine B. Candida albicans strains used in the experiment were mostly isolated from samples taken from patients suffering from variously located Candida albicans infections and four laboratory standard strains were also used. The method used for the experiment was a modified broth dilution method using combinations of both tyrosol and antimycotics various concentrations, which were prepared by two-fold dilution. Using amphotericine B, 28 strains did not show any difference in MIC compared with the control sample, 6 strains showed mildly higher MIC and 3 strains shower mildly lower MIC. Using fluconazole, 24...
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Detection of resistance to echinocandins antifungal agents in Candida sp. using molecular biological methods
Vitáčková, Petra ; Chrenková, Vanda (advisor) ; Nyč, Otakar (referee)
Invasive diseases due to Candida sp., especially by C. albicans, represent very severe complication in immunocompromised patients. More over the presence of antifungal resistance leads to delay of targeted antifungal therapy and increases morbidity and mortality in this group of patients. Therefore the aim was to introduce a rapid method of caspofungin resistance detection by the mass spectrometry MALDITOF. The tests were performed by the use of reference strain C. albicans CCM8261 and caspofungin resistant strain C. albicans M30. Different settings of mass spectrometer were used for the measuring. The obtained spectra were evaluated by correlation and cluster analysis (dendrogram). By cluster analysis it was possible to differenciate the susceptible and the resistant strain. During the analysis we have found, that mass spectrometer settings are unique for each machine and we cannot use the published data. We did not succeed to determine the similarity by correlation analysis. The quality of obtained spectra was quite poor, probably because of non-suitable culture medium used in the test The cluster analysis confirmed the possibility of resistance detection by mass spectrometry; nevertheless more testing profiting from current experience is needed for introduction of this test in routine. Powered by TCPDF...
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