Nejvíce citovaný článek - PubMed ID 19111673
Characterization of the kinetics and mechanisms of inhibition of drugs interacting with the S. cerevisiae multidrug resistance pumps Pdr5p and Snq2p
Saccharomyces species, which are mostly used in the food and beverage industries, are known to differ in their fermentation efficiency and tolerance of adverse fermentation conditions. However, the basis of their difference has not been fully elucidated, although their genomes have been sequenced and analyzed. Five strains of four Saccharomyces species (S. cerevisiae, S. kudriavzevii, S. bayanus, and S. paradoxus), when grown in parallel in laboratory conditions, exhibit very similar basic physiological parameters such as membrane potential, intracellular pH, and the degree to which they are able to quickly activate their Pma1 H+-ATPase upon glucose addition. On the other hand, they differ in their ability to proliferate in media with a very low concentration of potassium, in their osmotolerance and tolerance to toxic cations and cationic drugs in a growth-medium specific manner, and in their capacity to survive anhydrobiosis. Overall, S. cerevisiae (T73 more than FL100) and S. paradoxus are the most robust, and S. kudriavzevii the most sensitive species. Our results suggest that the difference in stress survival is based on their ability to quickly accommodate their cell size and metabolism to changing environmental conditions and to adjust their portfolio of available detoxifying transporters.
- Klíčová slova
- Intracellular pH, Membrane potential, Saccharomyces, Stress tolerance,
- MeSH
- fermentace MeSH
- fungální proteiny genetika metabolismus MeSH
- fyziologický stres MeSH
- glukosa metabolismus MeSH
- protonové ATPasy genetika metabolismus MeSH
- Saccharomyces klasifikace genetika růst a vývoj fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Názvy látek
- fungální proteiny MeSH
- glukosa MeSH
- protonové ATPasy MeSH
The maintenance of potassium homeostasis is crucial for all types of cells, including Candida glabrata. Three types of plasma-membrane systems mediating potassium influx with different transport mechanisms have been described in yeasts: the Trk1 uniporter, the Hak cation-proton symporter and the Acu ATPase. The C. glabrata genome contains only one gene encoding putative system for potassium uptake, the Trk1 uniporter. Therefore, its importance in maintaining adequate levels of intracellular potassium appears to be critical for C. glabrata cells. In this study, we first confirmed the potassium-uptake activity of the identified gene's product by heterologous expression in a suitable S. cerevisiae mutant, further we generated a corresponding deletion mutant in C. glabrata and analysed its phenotype in detail. The obtained results show a pleiotropic effect on the cell physiology when CgTRK1 is deleted, affecting not only the ability of trk1Δ to grow at low potassium concentrations, but also the tolerance to toxic alkali-metal cations and cationic drugs, as well as the membrane potential and intracellular pH. Taken together, our results find the sole potassium uptake system in C. glabrata cells to be a promising target in the search for its specific inhibitors and in developing new antifungal drugs.
- MeSH
- buněčná membrána metabolismus MeSH
- Candida glabrata metabolismus fyziologie MeSH
- draslík metabolismus MeSH
- homeostáza fyziologie MeSH
- iontový transport fyziologie MeSH
- kationty metabolismus MeSH
- membránové potenciály fyziologie MeSH
- proteiny přenášející kationty metabolismus MeSH
- regulace genové exprese u hub fyziologie MeSH
- Saccharomyces cerevisiae metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- draslík MeSH
- kationty MeSH
- proteiny přenášející kationty MeSH
- Trk1 protein, Candida albicans MeSH Prohlížeč
We investigated the impact of the deletions of genes from the final steps in the biosynthesis of ergosterol (ERG6, ERG2, ERG3, ERG5, ERG4) on the physiological function of the Saccharomyces cerevisiae plasma membrane by a combination of biological tests and the diS-C3(3) fluorescence assay. Most of the erg mutants were more sensitive than the wild type to salt stress or cationic drugs, their susceptibilities were proportional to the hyperpolarization of their plasma membranes. The different sterol composition of the plasma membrane played an important role in the short-term and long-term processes that accompanied the exposure of erg strains to a hyperosmotic stress (effect on cell size, pH homeostasis and survival of yeasts), as well as in the resistance of cells to antifungal drugs. The pleiotropic drug-sensitive phenotypes of erg strains were, to a large extent, a result of the reduced efficiency of the Pdr5 efflux pump, which was shown to be more sensitive to the sterol content of the plasma membrane than Snq2p. In summary, the erg4Δ and erg6Δ mutants exhibited the most compromised phenotypes. As Erg6p is not involved in the cholesterol biosynthetic pathway, it may become a target for a new generation of antifungal drugs.
- MeSH
- ABC transportéry genetika metabolismus MeSH
- antifungální látky farmakologie MeSH
- biosyntetické dráhy genetika MeSH
- buněčná membrána chemie fyziologie MeSH
- ergosterol biosyntéza chemie MeSH
- flukonazol farmakologie MeSH
- fluorescenční mikroskopie MeSH
- koncentrace vodíkových iontů MeSH
- membránové potenciály fyziologie MeSH
- methyltransferasy genetika metabolismus MeSH
- mnohočetná fungální léková rezistence účinky léků genetika fyziologie MeSH
- molekulární struktura MeSH
- mutace MeSH
- Saccharomyces cerevisiae - proteiny genetika metabolismus MeSH
- Saccharomyces cerevisiae chemie genetika fyziologie MeSH
- tolerance k soli genetika fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- ABC transportéry MeSH
- antifungální látky MeSH
- delta 24-sterol methyltransferase MeSH Prohlížeč
- ergosterol MeSH
- flukonazol MeSH
- methyltransferasy MeSH
- PDR5 protein, S cerevisiae MeSH Prohlížeč
- Saccharomyces cerevisiae - proteiny MeSH
- SNQ2 protein, S cerevisiae MeSH Prohlížeč
Three structurally related oxathiolone fused chalcone derivatives appeared effective chemosensitizers, able to restore in part sensitivity to fluconazole of multidrug-resistant C. albicans strains. Compound 21 effectively chemosensitized cells resistant due to the overexpression of the MDR1 gene, compound 6 reduced resistance of cells overexpressing the ABC-type drug transporters CDR1/CDR2 and derivative 18 partially reversed fluconazole resistance mediated by both types of yeast drug efflux pumps. The observed effect of sensitization of resistant strains of Candida albicans to fluconazole activity in the presence of active compounds most likely resulted from inhibition of the pump-mediated efflux, as was revealed by the results of studies involving the fluorescent probes, Nile Red, Rhodamine 6G and diS-C3(3).
- Klíčová slova
- Candida albicans, antifungals, chalcones, chemosensitization, multidrug resistance,
- Publikační typ
- časopisecké články MeSH
This review summarizes the main results obtained in the fields of general and molecular microbiology and microbial genetics at the Institute of Microbiology of the Academy of Sciences of the Czech Republic (AS CR) [formerly Czechoslovak Academy of Sciences (CAS)] over more than 50 years. Contribution of the founder of the Institute, academician Ivan Málek, to the introduction of these topics into the scientific program of the Institute of Microbiology and to further development of these studies is also included.
- MeSH
- akademie a ústavy dějiny MeSH
- dějiny 20. století MeSH
- mikrobiální genetika dějiny MeSH
- molekulární biologie dějiny MeSH
- Check Tag
- dějiny 20. století MeSH
- Publikační typ
- časopisecké články MeSH
- historické články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Geografické názvy
- Česká republika MeSH