The CYSTM (cysteine-rich transmembrane module) protein family comprises small molecular cysteine-rich tail-anchored membrane proteins found in many eukaryotes. The Saccharomyces cerevisiae strains carrying the CYSTM genes YDRO34W-B and YBR056W-A (MNC1) fused with GFP were used to test the expression of these genes under different stresses. The YBR056W-A (MNC1) and YDR034W-B genes are expressed under stress conditions caused by the toxic concentrations of heavy metal ions, such as manganese, cobalt, nickel, zinc, cuprum, and 2.4-dinitrophenol uncoupler. The expression level of YDR034W-B was higher than that of YBR056W-A under alkali and cadmium stresses. The Ydr034w-b-GFP and Ybr056w-a-GFP proteins differ in the cellular localization: Ydr034w-b-GFP was mainly observed in the plasma membrane and vacuolar membrane, while Ybr056w-a-GFP was observed in the cytoplasm, probably in intracellular membranes. The null-mutants in both genes demonstrated decreased cell concentration and lytic phenotype when cultivated in the presence of excess manganese. This allows for speculations about the involvement of Mnc1 and Ydr034w-b proteins in manganese stress overcoming.
This work was aimed to study of fungicidal activity and mechanism of action of the killer toxin of Cryptococcus pinus VKM Y-2958. A killer-toxin preparation was obtained and partly purified from the culture liquid of the yeast Cryptococcus pinus. Its fungicidal activity against the pathogenic yeast Filobasidiella neoformans was assessed by assay of target cells growth inhibition and propidium iodide staining. The preparation contained three peptides with molecular masses of about 11-13 kDa, which did not show similarities in the mass spectrometric analysis. When frozen, the killer-toxin preparation remained active for several years. The killer toxin of Cr. pinus exhibits the high fungicidal activity against the known pathogen F. neoformans, is stable during long-term storage, and its mechanism of action is based on damaging target cell membranes. The killer toxin was obtained by a simple procedure and is of interest for the development of novel fungicidal preparations.
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