Skip to main content
SpringerLink
Log in

Characterization of mutants of Escherichia coli with an increased control of translation fidelity

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

Summary

Neomycin-resistant mutants of Escherichia coli K12 were selected on a minimal succinate medium containing neomycin. In an in vitro poly(U)-dependent protein synthesizing assay, the ribosomes from the mutant strains showed lower levels of misreading than wild-type ribosomes in the presence of neomycin or other error-promoting aminoglycoside antibiotics or ethanol. The mutation was shown to affect a component of the 30S subunit and to facilitate the dissociation of ribosomes into subunits. It is suggested that an impairment in the subunit interaction may be responsible for the observed restriction of the stimulation of misreading.

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

ATP:

adenosine-5′-triphosphate

GTP:

guanosine-5′-triphosphate

poly(U):

polyuridylic acid

References

  • Apirion D, Schlessinger D (1968a) Coresistance to neomycin and kanamycin by mutations in an Escherichia coli locus that affects ribosomes. J Bacteriol 96:768–776

    Google Scholar 

  • Apirion D, Schlessinger D (1968b) Mapping and complementation of three genes specifying 30S ribosomal components in Escherichia coli. J Bacteriol 96:1431–1432

    Google Scholar 

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

    Google Scholar 

  • Buckel P, Buchberger A, Böck A, Wittmann HG (1977) Alteration of ribosomal protein L6 in mutants of Escherichia coli resistant to gentamicin. Mol Gen Genet 158:47–54

    Google Scholar 

  • Chang FN, Flaks JG (1972) Binding of dihydrostreptomycin to Escherichia coli ribosomes. Characteristics and equilibrium of the reaction. Antimicrob Agents Chemother 2:294–307

    Google Scholar 

  • Dabbs ER (1977) Correlation between a novel phenotype towards streptomycin and the binding of an additional protein to the ribosome in mutants of Escherichia coli B. Mol Gen Genet 158:55–61

    Google Scholar 

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

    Google Scholar 

  • Davies J, Davis BD (1968) Misreading of ribonucleic acid code words induced by aminoglycoside antibiotics. The effect of drug concentration. J Biol Chem 243:3312–3316

    Google Scholar 

  • Davis BD, Mingioli ES (1950) Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol 60:17–28

    Google Scholar 

  • Davis BD, Tai PC, Wallace BJ (1974) Complex interactions of antibiotics with the ribosome. In: Nomura M, Tissières A, Lengyel P (eds) Ribosomes. Cold Spring Harbor Laboratory, New York, pp 771–789

    Google Scholar 

  • Downie JA, Gibson F, Cox GB (1979) Membrane adenosine triphosphatases of prokaryotic cells. Ann Rev Biochem 48:103–131

    Google Scholar 

  • Gorini L (1974) Streptomycin and misreading of the genetic code. In: Nomura M, Tissières A, Lengyel P (eds) Ribosomes. Cold Spring Harbor Laboratory, New York, pp 791–803

    Google Scholar 

  • Grisé-Miron L, Noreau J, Melançon P, Brakier-Gingras L (1981) Comparison of the misreading induced by streptomycin and neomycin. Biochim Biophys Acta 656:103–110

    Google Scholar 

  • Grisé-Miron L, Brakier-Gingras L (1982) Effect of neomycin and protein S1 on the binding of streptomycin to the ribosome. Eur J Biochem 123:643–646

    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

    Google Scholar 

  • Knopf UC, Sommer A, Kenny J, Traut RR (1975) A new two-dimensional gel electrophoresis system for the analysis of complex protein mixtures. Application to the ribosome of E. coli. Mol Biol Reports 2:35–40

    Google Scholar 

  • Kühberger R, Piepersberg W, Petzer A, Buckel P, Böck A (1979) Alteration of ribosomal protein L6 in gentamicin-resistant strains of Escherichia coli. Effects on fidelity of protein synthesis. Biochemistry 18:187–193

    Google Scholar 

  • Lacoste L, Lacaille M, Brakier-Gingras L (1977) The role of misrepair processes in the isolation of new types of steptomycinresistant mutants in Escherichia coli. Mol Gen Genet 157:313–318

    Google Scholar 

  • Lando D, Cousin MA, Ojasoo T, Raynaud JP (1976) Paromomycin and dihydrostreptomycin binding to Escherichia coli ribosomes. Eur J Biochem 66:597–606

    Google Scholar 

  • Lennox ES (1955) Transduction of linked genetic characters of the host by bacteriophage P1. Virology 1:190–206

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, New York, pp 201–205

    Google Scholar 

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

    Google Scholar 

  • Nomura M, Morgan EA, Jaskunas SR (1977) Genetics of bacterial ribosomes. Ann Rev Genet 11:297–347

    Google Scholar 

  • Noreau J, Grisé-Miron L, Brakier-Gingras L (1980) Comparison of the action of streptomycin and neomycin on the structure of the bacterial ribosome. Biochim Biophys Acta 608:72–81

    Google Scholar 

  • Olsson MO, Isaksson LA (1979) Analysis of rps D mutations in Escherichia coli 1: Comparison of mutants with various alterations in ribosomal protein S4. Mol Gen Genet 169:251–257

    Google Scholar 

  • Piepersberg W, Geyl D, Hummel H, Böck A (1980) Physiology and biochemistry of bacterial ribosomal mutants. In: Osawa S, Ozeki H, Uchida H, Yura T (eds) Genetics and evolution of RNA polymerase, tRNA and ribosomes. University of Tokyo Press, Tokyo, pp 359–377

    Google Scholar 

  • Rosner JL (1972) Formation, induction and curing of bacteriophage P1 lysogens. Virology 49:679–689

    Google Scholar 

  • Stöffer G, Wittmann HG (1977) Primary structure and three-dimensional arrangement of proteins within the Escherichia coli ribosome. In: Weissbach H, Pestka S (eds) Molecular mechanism of protein synthesis. Academic Press, New York, pp 117–202

    Google Scholar 

  • Tai PC, Davis BD (1979) Triphasis concentration effects of gentamicin on activity and misreading in protein synthesis. Biochemistry 18:193–198

    Google Scholar 

  • Tanaka N (1975) Aminoglycoside antibiotics. In: Corcoran JW, Hahn FE (eds) Antibiotics III. Springer Verlag, New York, pp 340–364

    Google Scholar 

  • Thompson RC, Stone PJ (1977) Proofreading of the codon-antibodon interaction on ribosomes. Proc Natl Acad Sci USA 74:198–202

    Google Scholar 

  • Thompson RC, Dix DB, Gerson RB, Karim AM (1981) Effect of Mg2+ concentration, polyamines, streptomycin and mutations in ribosomal proteins on the accuracy of the two-step selection of aminoacyl-tRNAs in protein biosynthesis. J Biol Chem 256:6676–6681

    Google Scholar 

  • Thorbjarnardottir SH, Magnusdottir RA, Eggertsson G, Kagan SA, Andresson OS (1978) Mutations determining generalized resistance to aminoglycoside antibiotics in Escherichia coli. Mol Gen Genet 161:89–98

    Google Scholar 

  • Yarus M (1979) The accuracy of translation. Prog Nucleic Acid Res Mol Biol 23:195–225

    Google Scholar 

  • Yates JL (1979) Role of ribosomal protein S12 in discrimination of aminoacyl-tRNA. J Biol Chem 254:11550–11554

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Département de Biochimie, Université de Montréal, H3C 3J7, Montréal, Québec, Canada

    P. Phoenix, P. Melançon & L. Brakier-Gingras

Authors
  1. P. Phoenix
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. P. Melançon
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L. Brakier-Gingras
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by K. Isono

the symbols Nmr, Smr and Gmr denote resistance to neomycin, streptomycin and gentamicin, respectively

Rights and permissions

Reprints and permissions

About this article

Cite this article

Phoenix, P., Melançon, P. & Brakier-Gingras, L. Characterization of mutants of Escherichia coli with an increased control of translation fidelity. Molec Gen Genet 189, 123–128 (1983). https://doi.org/10.1007/BF00326064

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00326064

Keywords

Search

Navigation