Alterations in protein synthesis and levels of heat shock 70 proteins in response to salt stress of the halotolerant yeast Rhodotorula mucilaginosa

Responses of the halotolerant yeast Rhodotorula mucilaginosa YRH2 to salt stress was studied. Strain YRH2 was isolated from chemical industry park wastewater evaporation ponds that are characterized by large fluctuations in salinity and pH. Upon shift to high salt medium there is a shutdown of protein synthesis. Radiolabeling and separation of proteins from salt stressed and non-stressed cells identified down-regulated heat shock 70 proteins Ssb1/2p, by N-terminal sequencing and Western blotting. Ssb's role in salt stress in both R. mucilaginosa and S. cerevisiae was examined and we show that its response to salt stress and amino acid limitation is similar. Other proteins such as the heat shock 70 protein Kar2p/BiP and Protein Disulfide Isomerase were strongly induced in response to a shift to high salt in R. mucilaginosa and reacted in a manner similar to the effect of tunicamycin, a known unfolded protein response inducer. Also, assaying carboxypeptidase Y, we showed that high salt medium reduces the specific activity of the enzyme in R. mucilaginosa. It is suggested that the changes in the expression of the heat shock 70 proteins is a part of a mechanism which alleviates the damaging effects of high salt on protein folding in the yeast Rhodotorula mucilaginosa.     

Regulation of Mg,K-ATPase Activity in Guard Cells of Commelina communis by Phytochrome and ABA

The microsomal fraction obtained from guard cell protoplastswas assayed for ATPase activity at pH 6.5. Triton X-100 didnot affect this stimulation of activity of ATPase by K⁺ up to5 mM, but the detergent abolished the stimulatory effect ofK⁺ at higher concentrations. The ATPase activity was inhibitedby N,N-dicyclohexylcarbodiimide and ABA. Irradiation with redlight enhanced the ATPase activity more than did irradiationwith far-red light. ABA and irradiation with red or far-redlight were effective only in the presence of K⁺. These resultssuggest the possibility that the ATPase activity is modulatedonly indirectly by light and ABA. (Received January 9, 1989; Accepted July 10, 1989)