Acidic proteins of the large ribosomal subunit in Saccharomyces cerevisiae. Effect of phosphorylation.

Three strongly acidic proteins with pIs between 3.0 and 3.5 have been detected and purified from an ammonium-ethanol extract of Saccharomyces cerevisiae ribosomes. The three proteins, called L45, L44, and L44', have a similar amino acid composition, but differences were shown by tryptic peptide analysis. Nevertheless, the three polypeptides show total cross-reaction to antisera raised against one of them. Protein L44' is very unstable in the extract when treated at the basic pH 9.2, due to an enzymatic process not yet clarified. When purified, the protein is, however, stable. In solution, the proteins are present as dimers, as verified by ultracentrifugation, column filtration, and photochemical cross-linking. The tendency to dimerization is much lower in the case of protein L44'. On the average, 3.2 copies of these proteins are detected per ribosome. The proteins are monophosphorylated when present in the ribosome. Phosphorylation seems to regulate the affinity of the polypeptides for the particles because unphosphorylated proteins bind poorly to the ribosomes deprived of the acidic proteins. Since these proteins are unphosphorylated when present in the cytoplasm [Zinker, S. (1980) Biochim. Biophys. Acta 606, 76-82; Sánchez-Madrid, F., Vidales, F. J., & Ballesta, J. P. G. (1981) Eur. J. Biochem. 114, 609-613], a regulatory mechanism of the ribosomal function based on a phosphorylation-dephosphorylation process of the acidic proteins is being studied.