Abstract
The ribosomal stalk is involved directly in the interaction of the elongation factors with the ribosome during protein synthesis. The stalk is formed by a complex of five proteins, four small acidic polypepties and a larger protein which directly interacts with the rRNA at the GTPase center. In eukaryotes, the acidic components correspond to the 12 kDa P1 and P2 proteins, and the RNA binding component is protein P0. All these proteins are found to be phosphorylated in eukaryotic organisms. Previousin vitro data suggested this modification was involved in the activity of this structure. To confirm this possibility a mutational study has shown that phosphorylation takes place at a serine residue close to the carboxyl end of proteins P1, P2 and P0. This serine is part of a consensus casein kinase II phosphorylation site. However, by using a yeast strain carrying a temperature sensitive mutant, it has been shown that CKII is probably not the only enzyme responsible for this modification. Three new protein kinases, RAPI, RAPII and RAPIII, have been purified and compared with CKII and PK60, a previously reported enzyme that phosphorylates the stalk proteins. Differences among the five enzymes have been studied. It has also been found that some typical effects of the PKC kinase stimulate thein vitro phosphorylation of the stalk proteins. All the data available suggest that phosphorylation, although it is not involved in the interaction of the acidic proteins with the ribosome, affects ribosome activity and might participate in some ribosome regulatory mechanism.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
van Agthoven A., Kriek J., Amons R., Möller W.: Isolation and characterization of the acidic phosphoproteins of 60S ribosomes fromArtemia salina and rat liver.Eur. J. Biochem. 91, 553–556 (1978).
Amons R., Pluijms W., Möller W.: The primary structure of ribosomal protein eL12/eL12P fromArtemia salina 80S ribosomes.FEBS Lett. 104, 85–89 (1979).
Bailey-Serres J., Vangala S., Szick K., Lee C.H.: Acidic phosphoprotein complex of the 60S ribosomal subunit of maize seedling roots. Components and changes in response to flooding.Plant Physiol. 114, 1293–1305 (1997).
Ballesta J.P.G., Remacha M.: The large ribosomal subunit stalk as a regulatory element of the eukaryotic translational machinery.Progr. Nucl. Acad Res. Mol. Biol. 55, 157–193 (1996).
Hanna D.E., Rethinaswamy A., Glover C.V.: Casein kinase II is required for cell cycle progression during G1 and G2/M inSaccharomyces cerevisiae.J. Biol. Chem. 270, 25905–25914 (1991).
Hasler P., Brot N., Weisbach H., Parnassa A.P., Elkon K.B.: Ribosomal proteins P0, P1, and P2 are phosphorylated by casein kinase II at their conserved carboxyl termini.J. Biol. Chem. 266, 13815–13820 (1991).
Juan-Vidales F., Saenz-Robles M.T., Ballesta J.P.G.: Acidic proteins of the large ribosomal subunit inSaccharomyces cerevisiae. Effect of phosphorylation.Biochemistry 23, 390–396 (1984).
Kudlicki W., Szyszka R., Palen E., Gasior E.: Evidence for a highly specific protein kinase phosphorylating two strongly acidic proteins of yeast 60S ribosomal subunit.Biochim. Biophys. Acta 633, 376–385 (1980).
Kuenzel E.A., Mulligan J.A., Sommercorn J., Krebs E.G.: Substrate specificity determinants for casein kinase II as deduced from studies with synthetic peptides.J. Biol. Chem. 262, 9136–9140 (1987).
Lin A., Wittman-Leibold B., McNelly T., Wool I.G.: The primary structure of the acidic phosphoprotein from rat liver 60S ribosomal subunit.J. Biol. Chem. 257, 9189–9197 (1982).
Macconnell W.P., Kaplan N.O.: The activity of the acidic phosphoproteins from the 80S rat liver ribosome.J. Biol. Chem. 257, 5359–5366 (1982).
Pilecki M., Grankowski N., Jacobs J., Gąsior E.: Specific protein kinase fromSaccharomyces cerevisiae cells phosphorylating 60S ribosomal subunit.Eur. J. Biochem. 206, 259–267 (1992).
Remacha M., Jimenez-Diaz A. Bermejo B., Rodriguez-Gabriel M.A., Guarinos E., Ballesta J.P.G. Ribosomal acidic phosphoproteins P1 and P2 are not required for cell viability but regulate the pattern of protein expression inSaccharomyces cerevisiae.Mol. Cell. Biol. 15, 4754–4762 (1995).
Remacha M., Santos C., Ballesta J.P.G.: Disruption of single-copy genes encoding acidic ribosomal proteins inSaccharomyces cerevisiae.Mol. Cell. Biol. 10, 2182–2190 (1990).
Remacha M., Santos C., Bermejo B., Naranda T., Ballesta J.P.G.: Stable binding of the eukaryotic acidic phosphoproteins to the ribosome is not an absolute requirement forin vivo protein synthesis.J. Biol. Chem. 267, 12061–12067 (1992).
Rethinaswamy A., Birnbaum M.J., Glover C.V.: Temperature-sensitive mutations of theCKA1 gene reveal a role for casein kinase II in maintenance of cell polarity inSaccharomyces cerevisiae.J. Biol. Chem. 273, 5869–5877 (1998).
Sanchez-Madrid F., Juan-Vidales F., Ballesta J.P.G.: Effect of phosphorylation on affinity of acidic proteins fromSaccharomyces cerevisiae for the ribosome.Eur. J. Biochem. 114, 609–613 (1981).
Sanchez-Madrid F., Reyes R., Conde P., Ballesta J.P.G.: Acidic ribosomal proteins from eukaryotic cells. Effect on ribosomal functions.Eur. J. Biochem. 98, 409–416 (1979).
Sanchermann J., Krüger A., Kristiansen K.: Characterization of acidic proteins inTetrahymena pyriformis.FEBS Lett. 107, 343–347 (1979).
Santos C., Ballesta J.P.G.: Ribosomal phosphoprotein P0, contrary to phosphoproteins P1 and P2, is required forS. cerevisiae viability and ribosome assembly.J. Biol. Chem. 269, 15689–15696 (1994).
Santos C., Ballesta J.P.G.: The highly conserved protein P0 carboxyl end is essential for ribosome activity only in the absence of proteins P1 and P2.J. Biol. Chem. 270, 20608–20614 (1995).
Scharf K.-D., Nover L.: Control of ribosome biosynthesis in plant cell cultures under heat shoek conditions. II. Ribosomal proteins.Biochim. Biophys. Acta 909, 44–57 (1987).
Szyszka R., Bou G., Ballesta J.P.G.: RAP kinase, a new enzyme phosphorylating the acidic P proteins fromSaccharomyces cerevisiae ribosomes.Biochim. Biophys. Acta 1293, 213–221 (1996).
Tsurugi K., Ogata K.: Evidence for the exchangeability of acidic ribosomal proteins on cytoplasmic ribosomes in regenerating rat liver.J. Biochem. 98, 1427–1431 (1985).
Vard C., Guillot D., Bargis P., Lavergne J.P., Reboud J.P. A specific role for the phosphorylation of mammalian acidic ribosomal protein P2.J. Biol. Chem. 272, 20259–20262 (1997).
Vazquez M.P., Schijman A.G., Levin M.J.: Nucleotide sequence of a cDNA encoding aTryponosoma, cruzi acidic ribosomal P1 type protein.Nucl. Acids Res. 20, 2892 (1992a).
Vazquez M.P., Schuijman A.G., Panebra A., Levin M.J.: Nucleotide sequence of a cDNA encoding anotherTrypanosoma cruzi acidic ribosomal P2 type protein (TcP2b).Nucl. Acids Res. 20, 2893 (1992b).
Zambrano R., Briones E., Remacha M., Ballesta J.P.G.: Phosphorylation of the acidic ribosomal P proteins inSaccharomyces cerevisiae. A reappraisal.Biochemistry 36, 14439–1446 (1997).
Zinker S.: P5/P5′—the acidic ribosomal phosphoprotein fromS. cerevisiae.Biochim. Biophys. Acta 606, 76–82 (1980).
Zinker S., Warner J.R.: The ribosomal proteins ofSaccharomyces cerevisiae. Phosphorylated and exchangeable proteins.J. Biol. Chem. 251, 1799–1807 (1976).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Rodriguez-Gabriel, M.A., Bou, G., Briones, E. et al. Structure and function of the stalk, a putative regulatory element of the yeast ribosome. Role of stalk protein phosphorylation. Folia Microbiol 44, 153–163 (1999). https://doi.org/10.1007/BF02816234
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF02816234