Skip to main content
SpringerLink
Log in

Multiple effects of kanamycin on translational accuracy

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

Summary

We have studied the effects of kanamycin on the accuracy of translation in vitro by wild-type and mutant ribosomes from Escherichia coli. Kanamycin stimulates the leucine missense error of poly(U) translation by wild-type, Ram, and streptomycin-resistant ribosomes in characteristic ways; in particular, the streptomycin-resistant ribosomes are significantly less error-prone than wild-type or Ram ribosomes at all concentrations of the antibiotic. Kinetic analysis of the effects of kanamycin on the translational accuracy of wild-type ribosomes reveals a different concentration dependence for the perturbation of the initial selectivity and for the proofreading. Furthermore, the initial selectivity of streptomycin-resistant ribosomes is not affected by kanamycin; the drug enhances only the error of proofreading by this mutant ribosome. We suggest that the multiple effects of kanamycin on the errors of translation are due to separate effects at different ribosomal sites.

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

Abbreviations

N-AcPhe:

N-acetylphenylalanine

Km:

Kanamycin (used in the Figures and Tables only)

Str:

streptomycin (-′'-)

EF:

elongation factor

TCA:

trichloroacetic acid

Ram:

ribosome ambiguity mutant

References

  • Andersson DI, Kurland CG (1983) Ram ribosomes are defective proofreaders. Mol Gen Genet 191:378–381

    PubMed  Google Scholar 

  • Andersson DI, Bohman K, Isaksson LA, Kurland CG (1982) Translation rates and misreading characteristics of rpsD mutants in Escherichia coli. Mol Gen Genet 187:467–472

    PubMed  Google Scholar 

  • Arai KI, Kawakita M, Kaziro Y (1972) Studies on polypeptide elongation factors from Escherichia coli. J Biol Chem 247:7029–7037

    PubMed  Google Scholar 

  • Birge EA, Kurland CG (1969) Altered ribosomal protein in streptomycin-dependent Escherichia coli. Science 166:1282–1284

    PubMed  Google Scholar 

  • Cabanas MJ, Vazquez D, Modolell J (1978) Inhibition of ribosomal translocation by aminoglycoside antibiotics. Biochem Biophys Res Commun 83:991–997

    PubMed  Google Scholar 

  • Choi EC, Nishimura T, Tanaka N (1980) Mutational alterations of either large or small ribosomal subunit for the kanamycin resistance. Biochem Biophys Res Commun 94:755–762

    PubMed  Google Scholar 

  • Davies J, Davis BD (1968) Misreading of ribonucleic acid codewords induced by aminoglycoside antibiotics. J Biol Chem 243:3312–3316

    PubMed  Google Scholar 

  • Davies J, Gilbert W, Gorini L (1964) Streptomycin, suppression, and the code. Proc Natl Acad Sci USA 51:883–890

    PubMed  Google Scholar 

  • Davies J, Gorini L, Davis BD (1965) Misreading of RNA codewords induced by aminoglycoside antibiotics. Mol Pharmacol 1:93–106

    PubMed  Google Scholar 

  • Hopfield JJ (1974) Kinetic Proofreading: a new mechanism for reducing errors in biosynthetic processes requiring high specificity. Proc Natl Acad Sci USA 71:4135–4139

    PubMed  Google Scholar 

  • Jelenc PC (1980) Rapid purification of highly active ribosomes from Escherichia coli. Anal Biochem 105:369–374

    PubMed  Google Scholar 

  • Jelenc PC, Kurland CG (1979) Nucleoside triphosphate regeneration decreases the frequency of translation errors. Proc Natl Acad Sci USA 76:3174–3178

    PubMed  Google Scholar 

  • Kurland CG, Andersson DI, Andersson SGE, Bohman K, Bouadloun F, Ehrenberg M, Jelenc PC, Ruusala T (1983) Translational accuracy and bacterial growth. Alfred Benzon Symposium 19, Gene Expression. Munksgaard, Copenhagen, in press

    Google Scholar 

  • Leberman R, Antonsson B, Giovanelli R, Guarigata R, Schuman R, Wittinghofer A (1980) A simplified procedure for the isolation of bacterial polypeptide elongation factor EF-Tu. Anal Biochem 104:29–36

    PubMed  Google Scholar 

  • Misumi M, Tanaka N (1980) Mechanism of inhibition of translocation by kanamycin and viomycin: a comparative study with fusidic acid. Biochem Biophys Res Commun 92:647–654

    PubMed  Google Scholar 

  • Misumi M, Nishimura T, Komai T, Tanaka N (1978) Interaction of kanamycin and related antibiotics with the large subunit of ribosomes and the inhibition of translocation. Biochem Biophys Res Commun 84:358–365

    PubMed  Google Scholar 

  • Ninio J (1975) Kinetic amplification of enzyme discrimination. Biochimie 57:587–595

    PubMed  Google Scholar 

  • Olsson MO, Isaksson LA (1979) Analysis of rpsD mutations in Escherichia coli I: comparison of mutants with various alterations in ribosomal protein S4. Mol Gen Genet 169:251–257

    PubMed  Google Scholar 

  • Okuyama A, Tanaka N (1972) Differential effects of aminoglycosides on cistron-specific initiation of protein synthesis. Biochem Biophys Res Commun 49:951–957

    PubMed  Google Scholar 

  • Ozaki M, Mizushima S, Nomura M (1969) Identification and functional characterization of the protein controlled by the streptomycin-resistant locus in Escherichia coli. Nature 222:333–339

    PubMed  Google Scholar 

  • Ruusala T, Ehrenberg M, Kurland CG (1982) Is there proofreading during polypeptide synthesis? EMBO J 1:741–745

    PubMed  Google Scholar 

  • Semenkov YuP, Katunin VI, Makarov EM, Kirillov SV (1982) Quantitative study of kanamycin action on different functions of Escherichia coli ribosomes. FEBS Lett 144:121–124

    Article  Google Scholar 

  • Vazquez D (1979) Inhibitors of protein synthesis, Springer-Verlag, Berlin

    Google Scholar 

  • Wagner EGH, Kurland CG (1980) Escherichia coli elongation factor G blocks stringent factor. Biochemistry 19:1234–1240

    PubMed  Google Scholar 

  • Wanger EGH, Jelenc PC, Ehrenberg M, Kurland CG (1982) Rate of elongation of polyphenylalanine in vitro. Eur J Biochem 122:193–197

    PubMed  Google Scholar 

  • Zierhut G, Piepersberg W, Böck A (1979) Comparative analysis of the effects of aminoglycosides on bacterial protein synthesis in vitro. Eur J Biochem 98:577–583

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Molecular Biology, Biomedicum, Box 590, S-751 24, Uppsala, Sweden

    Pierre C. Jelenc & C. G. Kurland

Authors
  1. Pierre C. Jelenc
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C. G. Kurland
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by A. Böck

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jelenc, P.C., Kurland, C.G. Multiple effects of kanamycin on translational accuracy. Molec. Gen. Genet. 194, 195–199 (1984). https://doi.org/10.1007/BF00383516

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00383516

Keywords

Search

Navigation