Summary
The number of specific binding sites for homogenous single ribosomal proteins on 16S E. coli ribosomal RNA was investigated. The capacity of each of the twenty-one 30S subunit proteins to bind to the RNA was estimated by two newly developed methods, namely immunoprecipitation and a polyacrylamide gel method. Five proteins, namely S4, S7, S8, S15 and S20 bound specifically. One, S17, bound nonspecifically. No binding of the other proteins was detected. The binding proteins bound simultaneously to the RNA, with stimulated binding of proteins S7 and S8. Evidence is provided for the similarity of the chemistry of the binding sites of the binding proteins in Escherichia coli and in Bacillus stearothermophilus ribosomes.
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Chrambach, A., Reisefeld, R. A., Wyckoff, M., Zaccari, J.: A procedure for rapid and sensitive staining of protein fractionated by polyacrylamide gel electrophoresis. Analyt. Biochem. 20, 150–154 (1967).
Craven, G. R., Voynow, P., Hardy, S. J. S., Kurland, C. G.: The ribosomal proteins of Escherichia coli II. Chemical and physical characterisation of the 30S ribosomal proteins. Biochemistry 8, 2906–2915 (1969).
Dzionara, M., Kaltschmidt, E., Wittmann, H. G.: Ribosomal proteins. XIII: Molecular weights of isolated ribosomal proteins of Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 67, 1909–1913 (1970).
Gesteland, R. F.: Isolation and characterisation of ribonuclease I mutants of Escherichia coli: J. molec. Biol. 16, 67–84 (1966).
Hardy, S. J. S., Kurland, C. G., Voygnow, P., Mora, G.: The ribosomal proteins of Escherichia coli. I. Purification of the 30S ribosomal proteins. Biochemistry 8, 2897–2905 (1969).
Hastings, J. R. B. Parish, J. H., Kirby, K. S., Klucis, E.: Use of the zonal centrifuge to separate components of ribosomal RNA. Nat. Cancer Inst. Monogr. No. 21, 397–402 (1966).
Hill, W. E., Rossetti, G. P., Holde, K. E., van: Physical studies on ribosomes from Escherichia coli. J. molec. Biol. 44, 263–277 (1969).
Hiller, A., Plazin, A., Slyke, D. D. van: A study of conditions for Kjeldahl determination of nitrogen in proteins. J. biol. Chem. 176, 1401–1420 (1948).
Hindennach, I., Stöffler, G., Wittmann, H. G.: Ribosomal proteins IX. Isolation of proteins from 30S ribosomal subunits of Escherichia coli. Europ. J. Biochem. 23, 7–11 (1971).
Kaltschmidt, E., Wittmann, H. G.: Ribosomal proteins VII. Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Analyt. Biochem 36, 401–412 (1970).
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin-phenol reagent. J. biol. Chem. 193, 265–275 (1951).
Meybaum, W.: Über die Bestimmung kleiner pentose Mengen insbesondere in derivaten der Adenylsäure. Hoppe-Seylers Z. physiol. Chem. 258 117–120 (1939).
Mizushima, S., Nomura, M.: Assembly mapping of 30S ribosomal proteins from Escherichia coli. Nature (Lond.) 226, 1214–1218 (1970).
Nomura, M., Traub, P., Bechmann, H.: Hybrid 30S ribosomal particles reconstituted from components of different bacterial origins. Nature (Lond.) 219, 793–799 (1968).
Ouchterlony, O.: Diffusion in gel. Methods for immunological analyses. Prog. Allergy 5, 1–78 (1958).
Robinson, H. K., Wade, H. E.: The preparation of ribosomal ribonucleic acid from whole bacteria. Biochem. J. 106, 897–903 (1968).
Salas, M., Smith, M. A., Stanley, W. M., Wahba, A. J., Ochoa, S.: Direction of reading of the genetic message. J. biol. Chem. 240, 3988–3995 (1965).
Schaup, H. W., Green, M., Kurland, C. G.: Molecular interactions of ribosomal components. I. Identification of RNA binding sites for individual 30S ribosomal proteins. Molec. gen. Genet. 109, 193–205 (1970).
—: Molecular interactions of ribosomal components. II. Site-specific complex formation between 30S proteins and ribosomal RNA. Molec. gen. Genet. 112, 1–8 (1971).
Stöffler, G.: A rapid micromethod for electrophoretic separation of Escherichia coli ribosomal proteins on gelatinized cellulose acetate. Molec. gen. Genet. 100, 374–377 (1967).
—: Immunologische Untersuchung mit Antiseren gegen isolierte ribosomale Proteine aus 30S und 50S Untereinheiten aus Escherichia coli. Hoppe-Seylers Z. physiol. Chem. 350, 116 (1969).
—, Wittmann, H. G.: Ribosomal proteins XXIII. Sequence differences of Escherichia coli 30S ribosomal proteins as determined by immunochemical methods. Proc. nat. Acad. Sci. (Wash.) 68, 2283–2287 (1971).
Traub, P., Nomura, M.: Structure and function of Escherichia coli ribosomes. VI. Mechanism of assembly of 30S ribosomes studied in vitro. J. molec. Biol. 40, 391–413 (1969).
Wittmann, H. G., Stöffler, G., Hindennach, I., Kurland, C. G., Randall-Hazelbauer, L., Birge, E. A., Nomura, M., Kaltschmidt, E. Mizushima, S., Traut, R. R., Bickle, T. A.: Correlation of 30S ribosomal proteins of Escherichia coli isolated in different laboratories. Molec. gen. Genet. 111, 327–333 (1971).
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Communicated by P. Starlinger
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Garrett, R.A., Rak, K.H., Daya, L. et al. Ribosomal proteins. Molec. Gen. Genetics 114, 112–124 (1972). https://doi.org/10.1007/BF00332782
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DOI: https://doi.org/10.1007/BF00332782