Purification and properties of a double‐stranded ribonuclease from the yeast Saccharomyces cerevisiae

Publisher Website
Google Scholar
Abstract
A double‐stranded ribonuclease has been purified more than 90‐fold to near homogeneity from the yeast, Saccharomyces cerevisiae. The enzyme shows a high specificity for double‐stranded RNA as its substrate. It has a molecular weight of 27000 as determined by sodium dodecyl sulphate/polyacrylamide gel electrophoresis. The enzyme degrades dsRNA optimally at 30°C; it is stimulated by KCl and by the –SH reagent, dithiothreitol. In contrast to RNase III from Escherichia coli, the yeast enzyme is inhibited by divalent cations. Physiological studies have demonstrated that in vivo levels of the enzyme activity fell during the latter part of the exponential growth phase but rose during stationary phase. The specific activity of the enzyme in nitrogen‐starved yeast cells was 2–3‐fold higher than in non‐starved cells. The enzyme could be detected in yeast strains containing both, one or none of the species of cytoplasmic dsRNA (L and MdsRNAs) and may, therefore, have some wider role.