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Characterisation of the ribosomal proteins from Schizosaccharomyces pombe by two-dimensional polyacrylamide gel electrophoresis

Demonstration that a cycloheximide resistant strain, cyh1, has an Altered 60S ribosomal protein

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The 80S ribosome of Schizosaccharomyces pombe contains 93 proteins as determined by twodimensional electrophoresis on polyacrylamide gels. Of these, 76 are basic and 17 are acidic at pH 8.7. 38 proteins could be assigned unambiguosly to the large sub-unit and 19 to the small.

67 proteins were extracted from the two-dimensional gels and their molecular weights determined by electrophoresis on calibrated SDS-gels. Values varied from 11,000 to 52,000 daltons, the number average being 25,000 daltons. Hence the total protein content of the 80S ribosome must be at least 1.67x106 daltons.

Three drug resistant strains are known, cyh1, anil and tri5 (resistant to cycloheximide, anisomycin and trichodermin respectively), in which resistace is conferred by an altered ribosome, in each case by an altered 60S sub-unit. When 80S ribosomal protein patterns from these strains were compared with that of wild type, in only one case was a clear difference seen. This involved a large sub-unit, basic protein (designated number 66 on our classification) which, in the cyh1 strain, had a reduced mobility in the second dimension when compared to the wild type. The mutant form of protein 66 had a molecular weight of 25,000 daltons compared to the 22,000 of the wild type protein. Production of a larger protein by the mutant strain could either be due to a readthrough event or to an alteration in the specificity of a modifying or processing enzyme.

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  1. Grant, P., Sanchez, L., Jimenez, A.: Cryptopleurine resistance: Genetic locus for a 40S ribosomal component in Saccharomyces cerevisiae. J. Bact. 120, 1308–1314 (1974)

  2. Gutz, H., Heslot, H., Leupold, U., Loprieno, N.: Schizosaccharomyces pombe. In: Handbook of genetics (ed. R.C. King), Vol. 1, pp. 395–446. New York and London: Plenum Press 1974

  3. Howard, G.A., Trough, J.A., Croser, E.A., Traut, R.R.: Ribosomal proteins from rabbit reticulocytes: Number and molecular weights of proteins from ribosomal subunits. J. molec. Biol. 93, 391–404 (1975)

  4. Ibrahim, M.A.K.: Isolation and characterisation of mutants of Schizosaccharomyces pombe with impaired ribosomal function. Ph. D. Thesis. University of East Anglia, Norwich (1976)

  5. Ibrahim, M.A.K., Coddington, A.: Genetic studies on cycloheximide resistant strains of Schizosaccharomyces pombe. Heredity 37, 179–191 (1976)

  6. Ishiguro, J.: Studies on proteins from yeast cytoplasmic ribosomes by two-dimensional gel electrophoresis. Molec. gen. Genet. 145, 73–79 (1976)

  7. Jimenez, A., Littlewood, B., Davies, J.: Inhibition of protein synthesis in yeast. In: Molecular mechanisms of antibiotic action on protein synthesis and membranes (eds. E. Munoz, F. Garcia-Fernandez and D. Vazquez), pp. 292–306. Amsterdam: Elsevier 1972

  8. Kaltschmidt, E., Wittmann, H.G.: Ribosomal Proteins. VII. Twodimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Anal. Biochem. 36, 401–412 (1970)

  9. Mazelis, A.G., Peterman, M.L.: Physical-chemical properties of stable yeast ribosomes and ribosomal subunits. Biochim. biophys. Acta (Amst.) 312, 111–121 (1973)

  10. Schindler, D., Grant, P., Davies, J.: Trichodermin resistance, a mutation affecting eukaryotic ribosomes. Nature (Lond.) 248, 535–536 (1974)

  11. Sherton, C.C., Wool, I.G.: The extraction of proteins from eukaryotic ribosomes and ribosomal subunits. Molec. gen. Genet. 135, 97–112 (1974)

  12. Stöffler, G., Wool, I.G., Lin, A., Pak, K.H.: The identification of the eukaryotic ribosomal proteins homologous with Escherichia coli proteins L7 and L12. Proc. nat. Acad. Sci. (Wash.) 71, 4723–4726 (1974)

  13. Warner, J.R.: The assembly of ribosomes in yeast. J. biol. Chem. 246, 447–454 (1971)

  14. Weber, K., Osborn, M.: The reliability of molecular weight determination by dodecylsulphate polyacrylamide gel electrophoresis. J. biol. Chem. 244, 4406–4412 (1969)

  15. Welfle, H., Stahl, J., Bielka, H.: Studies on proteins of animal ribosomes. XIII. Enumeration of ribosomal proteins of rat liver. FEBS Letters 26, 228–232 (1972)

  16. Wittmann, H.G., Wittmann-Liebold, B.: Chemical structure of bacterial ribosomal proteins. In: Ribosomes (M. Normura, A. Tissieres and P. Lengyel, eds.), pp. 115–140. New York: Cold Spring Harbor Laboratory, 1974

  17. Zinker, S., Warner, J.R.: The ribosomal proteins of Saccharomyces cerevisiae. Phosphorylated and exchangeable proteins. J. biol. Chem. 251, 1799–1807 (1976)

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Correspondence to Alan Coddington.

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Communicated by F. Kaudewitz

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Coddington, A., Fluri, R. Characterisation of the ribosomal proteins from Schizosaccharomyces pombe by two-dimensional polyacrylamide gel electrophoresis. Molec. Gen. Genet. 158, 93–100 (1977). https://doi.org/10.1007/BF00455123

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  • Ribosomal Protein
  • Anil
  • Cycloheximide
  • Total Protein Content
  • Protein Pattern