Skip to main content
SpringerLink
Log in

Biochemical and genetic studies on two different types of erythromycin resistant mutants of Escherichia coli with altered ribosomal proteins

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Ribosomes from nine E. coli mutants with high level resistance to the antibiotic erythromycin were isolated and their proteins were compared with those of the parental strains by two-dimensional polyacrylamide gel electrophoresis, by carboxymethylcellulose column chromatography and by immunological techniques. Two 50S proteins were found to be altered in the mutants: either L 4 or L 22.

Ribosomes with an altered L4 protein bound erythromycin rather poorly and the formation of N-acetylphenylalanyl puromycin was drastically decreased. On the other handribosomes with an altered L22 protein bound erythromycin as efficiently as wild type ribosomes and their puromycin reaction was at least as good as that of wild type ribosomes.

Transduction experiments showed that the mutations affecting both proteins, L4 and L22, are located very close to the str and spc genes, nearer to the spc than to str gene.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Apirion, D.: Altered ribosomes in a suppressor strain of Escherichia coli. J. molec. Biol. 16, 285–301 (1966)

    Google Scholar 

  • Apirion, D.: Three genes that affect Escherichia coli ribosomes. J. molec. Biol. 30, 255–275 (1967)

    Google Scholar 

  • Apirion, D., Schlessinger, D.: Functional interdependence of ribosomal components of Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 63, 794–799 (1969)

    Google Scholar 

  • Dekio, S.: Genetic studies of ribosomal proteins in Escherichia coli. VII. Mapping of several ribosomal protein components by transduction experiments between Shigella dysenteriae and Escherichia coli and between different strains of Escherichia coli. Molec. gen. Genet. 113, 20–30 (1971)

    Google Scholar 

  • Dekio, S., Takata, R.: Genetic studies of the ribosomal proteins in Escherichia coli II. Altered 30S ribosomal protein component specific to spectinomycin resitant mutants. Molec gen. Genet. 105, 219–224 (1969)

    Google Scholar 

  • DeWilde, M., Wittmann-Liebold, B.: Localization of the amino acid exchanges in protein S5 from an Escherichia coli mutant resistant to spectinomycin. Molec. gen. gen. (1973) in press

  • Funatsu, G., Nierhaus, K., Wittmann, H. G.: Ribosomal proteins, XXXVII. Determination of allele types and amino acid exchanges in protein S12 of three streptomycin-resistant mutants of Escherichia coli. Biochim. biophys. Acta (Amst.) 287, 282–291 (1972b)

    Google Scholar 

  • Funatsu, G., Nierhaus, K., Wittmann-Liebold, B.: Ribosomal proteins. XXII. Studies on the altered protein S5 from a spectinomycin resistant mutant of Escherichia coli. J. molec. Biol. 64, 201–209 (1972a)

    Google Scholar 

  • Funatsu, G., Schiltz, E., Wittmann, H. G.: Ribosomal proteins XXVII. Localization of the amino acid exchanges in protein S5 from two Escherichia coli mutants resistant to spectinomycin. Molec. gen. Genet. 114, 106–111 (1971)

    Google Scholar 

  • Funatsu, G., Wittmann, H. G.: Ribosomal proteins XXXIII. Location of amino acid replacements in protein S12 isolated from Escherichia coli mutants resistant to streptomycin. J. molec. Biol. 68, 547–550 (1972)

    Google Scholar 

  • Gesteland, R. F.: Isolation and characterization of ribonuclease I mutants of Escherichia coli. J. molec. Biol. 16, 67–84 (1966)

    Google Scholar 

  • Hahn, F. E.: Erythromycin and oleandomycin. In: Antibiotics I. Mechanism of action, p. 378–386 (ed. by D. Gottlieb and P. D. Shaw). Berlin-Heidelberg-New York: Springer 1967

    Google Scholar 

  • Kaji, H., Tanaka, Y.: Binding of dihydrostreptomycin to ribosomal subunits. J. molec. Biol. 32, 221–230 (1968)

    Google Scholar 

  • Kaltschmidt, E., Wittmann, H. G.: Ribosomal proteins, VII. Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Analyt. Biochem. 36, 401–412 (1970b)

    Google Scholar 

  • Kaltschmidt, E., Wittmann, H. G.: Ribosomal proteins, XII. Number of proteins in small and large ribosomal subunits of Escherichia coli as determined by two-dimensional gel electrophoresis. Proc. nat. Acad. Sci. (Wash.) 67, 1276–1281 (1970a)

    Google Scholar 

  • Kaneda, T., Butte, J., Taubman, S. B., Corcoran, J. W.: Actinomycete antibiotics. III. The biogenesis of erythronolide, the C21 branched chain lactone in erythromycin. J. biol. Chem. 237, 322–328 (1962)

    Google Scholar 

  • Krembel, J., Apirion, A.: Changes in ribosomal proteins associated with mutants in a locus that affects Escherichia coli ribosomes. J. molec. Biol. 33, 363–368 (1968)

    Google Scholar 

  • Lapidot, Y., DeGroot, N., Fry-Shafir, I.: Modified aminoacyl-tRNA. II. A general method for the preparation of acylaminoacyl-tRNA. Biochim. biophys. Acta (Amst.) 145, 292–299 (1967)

    Google Scholar 

  • Lennette, E. T., Apirion, D.: Genetic analysis of an Escherichia coli syndrome. J. Bact. 108, 1322–1328 (1971)

    Google Scholar 

  • Nomura, M., Engbaek, F.: Expression of ribosomal protein genes as analyzed by bacteriophage Mu-induced mutations. Proc. nat. Acad. Sci. (Wash.) 69, 1526–1530 (1972)

    Google Scholar 

  • Osawa, S., Otaka, E., Itoh, T., Fukui, T.: Biosynthesis of 50S ribosomal subunit in Escherichia coli. J. molec. Biol. 40, 321–351 (1969)

    Google Scholar 

  • Osawa, S., Otaka, E., Takata, R., Dekio, S., Matsubara, K., Itoh, T., Muto, A., Tanaka, K., Teraoka, H., Tamaki, M.: Ribosomal protein genes in bacteria. FEBS Symp. vol. 23, p. 313–336. London: Academic Press 1972

    Google Scholar 

  • Osawa, S., Takata, R., Tanaka, K., Tamaki, M.: Chloramphenicol resistant mutants in Bacillus subtilis. Molec. gen. Genet. 127, 168–173 (1973)

    Google Scholar 

  • Otaka, E., Itoh, T., Osawa, S., Tanaka, K., Tamaki, M.: Peptide analyses of a protein component, 50-8, of 50S ribosomal subunit from erythromycin resistant mutants of Escherichia coli and Escherichia freundii. Molec. gen. Genet. 114, 14–22 (1971)

    Google Scholar 

  • Otaka, E., Teraoka, H., Tamaki, M., Tanaka, K., Osawa, S.: Ribosomes from erythromycinresistant mutants of Escherichia coli Q13. J. molec. Biol. 48, 499–510 (1970)

    Google Scholar 

  • Ozaki, M., Mizushima, S., Nomura, M.: Identification and functional characterization of the protein controlled by streptomycin resistant locus of E. coli. Nature (Lond.) 222, 333 (1969)

    Google Scholar 

  • Pestka, S.: Studies on the formation of transfer ribonucleic acid-ribosome complex. III. The formation of peptide bonds by ribosomes in the absence of supernatant enzymes. J. biol. Chem. 243, 2810–2820 (1968)

    Google Scholar 

  • Schreiner, G., Nierhaus, K.: Proteins involved in the binding of dehydrostreptomycin to ribosomes of Escherichia coli. J. molec. Biol., in press

  • Stöffler, G., Tischendorf, G., Hasenbank, R., Wittmann, H. G.: The number of individual proteins of the 50S ribosomal subunit from Escherichia coli as determined by immunochemical methods. Manuscript submitted

  • Stöffler, G., 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)

    Google Scholar 

  • Takata, R.: Genetic studies of the ribosomal proteins in Escherichia coli. VIII. Mapping of ribosomal protein components by intergeneric mating experiments between Serratia marcescens and Escherichia coli. Molec. gen. Genet. 118, 363–371 (1972)

    Google Scholar 

  • Takata, R., Osawa, S., Tanaka, K., Teraoka, H., Tamaki, M.: Genetic studies of the ribosomal proteins in Escherichia coli. V. Mapping of erythromycin resistance mutations which lead to alteration of a 50S ribosomal protein component. Molec. gen. Genet. 109, 123–130 (1970)

    Google Scholar 

  • Tanaka, K., Tamaki, M., Osawa, S., Kimura, A., Takata, R.: Erythromycin resistant mutants of Bacillus subtilis. Molec. gen. Genet. 127, 157–161 (1973)

    Google Scholar 

  • Teraoka, H.: A reversible change in the ability of Escherichia coli ribosomes to bind to erythromycin. J. molec. Biol. 48, 511–515 (1970)

    Google Scholar 

  • Teraoka, H., Tanaka, K.: Reaction of puromycin with N-acetyl-phenyl-alanyl-tRNA on ribosomes reassociated from Escherichia coli ribosomal subunits. Biochim. biophys. Acta (Amst.) 247, 304–309 (1971)

    Google Scholar 

  • Wheatherford, S. C., Rosen, L., Gorelic, L., Apirion, D.: Escherichia coli strains with thermolabile ribonuclease II activity. J. biol. Chem. 247, 5404–5408 (1972)

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Max-Planck-Institut für Molekulare Genetik, Berlin-Dahlem, Germany

    H. G. Wittmann & G. Stöffler

  2. Department of Microbiology, Washington University Medical School, St. Louis, USA

    D. Apirion & L. Rosen

  3. Shionogi Research Laboratory, Shionogi & Co. Ltd., Fukushima-ku, Osaka, Japan

    K. Tanaka & M. Tamaki

  4. Research Institute for Nuclear Medicine and Biology, University of Hiroshima, Hiroshima, Japan

    R. Takata, S. Dekio, E. Otaka & S. Osawa

Authors
  1. H. G. Wittmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. Stöffler
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. D. Apirion
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. L. Rosen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. K. Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. M. Tamaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. R. Takata
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. S. Dekio
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. E. Otaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. S. Osawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Paper No. 61 on “Ribosomal Proteins”. Preceding paper is by Hasenbank et al., Molec. gen. Genet., 127, 1–18 (1973).

Communicated by E. Bautz

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wittmann, H.G., Stöffler, G., Apirion, D. et al. Biochemical and genetic studies on two different types of erythromycin resistant mutants of Escherichia coli with altered ribosomal proteins. Molec. Gen. Genet. 127, 175–189 (1973). https://doi.org/10.1007/BF00333665

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00333665

Keywords

Search

Navigation