K2 killer toxin-induced physiological changes in the yeast Saccharomyces cerevisiae.

Saccharomyces cerevisiae cells produce killer toxins, such as K1, K2 and K28, that can modulate the growth of other yeasts giving advantage for the killer strains. Here we focused on the physiological changes induced by K2 toxin on a non-toxin-producing yeast strain as well as K1, K2 and K28 killer strains. Potentiometric measurements were adjusted to observe that K2 toxin immediately acts on the sensitive cells leading to membrane permeability. This correlated with reduced respiration activity, lowered intracellular ATP content and decrease in cell viability. However, we did not detect any significant ATP leakage from the cells treated by killer toxin K2. Strains producing heterologous toxins K1 and K28 were less sensitive to K2 than the non-toxin producing one suggesting partial cross-protection between the different killer systems. This phenomenon may be connected to the observed differences in respiratory activities of the killer strains and the non-toxin-producing strain at low pH. This might also have practical consequences in wine industry; both as beneficial ones in controlling contaminating yeasts and non-beneficial ones causing sluggish fermentation.

Regulation of the retention factor for weak acids in micellar electrokinetic chromatography with cationic surfactant via variation of the chloride concentration.

For tetradecyltrimethylammonium bromide in boric acid/borate or acetic acid/acetate buffer and NaCl or CaCl2 as the added salt, it is investigated whether the retention behaviour of weak acids in MEKC with cationic surfactant can be modelled by assuming for the deprotonated species predominantly electrostatic interaction with the micelles acting as a pseudostationary ion-exchanger. The retention of (partially) charged solutes by oppositely charged micelles is analyzed by applying the classical theory of IEC (plotting lg k against lg(c(Cl⁻)) assuming a fixed concentration of ion-exchange sites. When plotting the absolute slopes of the regression lines against the absolute effective charge numbers of the analytes, correlation coefficients of 0.968-0.998 were obtained. It is shown that the dependence of the retention factor on the concentration of chloride (the competing ion) in the separation electrolyte and on the degree of dissociation of the analyte corresponds to what would be expected for mixed-mode retention (hydrophobic and ion-exchange interaction) on a pseudostationary ion-exchanger.

Screening the budding yeast genome reveals unique factors affecting K2 toxin susceptibility.

BACKGROUND: Understanding how biotoxins kill cells is of prime importance in biomedicine and the food industry. The budding yeast (S. cerevisiae) killers serve as a convenient model to study the activity of biotoxins consistently supplying with significant insights into the basic mechanisms of virus-host cell interactions and toxin entry into eukaryotic target cells. K1 and K2 toxins are active at the cell wall, leading to the disruption of the plasma membrane and subsequent cell death by ion leakage. K28 toxin is active in the cell nucleus, blocking DNA synthesis and cell cycle progression, thereby triggering apoptosis. Genome-wide screens in the budding yeast S. cerevisiae identified several hundred effectors of K1 and K28 toxins. Surprisingly, no such screen had been performed for K2 toxin, the most frequent killer toxin among industrial budding yeasts. PRINCIPAL FINDINGS: We conducted several concurrent genome-wide screens in S. cerevisiae and identified 332 novel K2 toxin effectors. The effectors involved in K2 resistance and hypersensitivity largely map in distinct cellular pathways, including cell wall and plasma membrane structure/biogenesis and mitochondrial function for K2 resistance, and cell wall stress signaling and ion/pH homeostasis for K2 hypersensitivity. 70% of K2 effectors are different from those involved in K1 or K28 susceptibility. SIGNIFICANCE: Our work demonstrates that despite the fact that K1 and K2 toxins share some aspects of their killing strategies, they largely rely on different sets of effectors. Since the vast majority of the host factors identified here is exclusively active towards K2, we conclude that cells have acquired a specific K2 toxin effectors set. Our work thus indicates that K1 and K2 have elaborated different biological pathways and provides a first step towards the detailed characterization of K2 mode of action.