Skip to main content
Log in

Localized mutagenesis of the aroE-strA section of the Escherichia coli chromosome coding for ribosomal proteins

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

Summary

In order to obtain E. coli strains altered in ribosomal proteins the following isolation technique was used: Phage P1 grown in a streptomycin resistant E. coli strain, was mutagenized by hydroxylamine or nitrous acid, and was used to transduce into a strain auxotrophic for aroE. Transductants with streptomycin resistance and aroE prototrophy were selected and tested for their growth at various temperatures (20°, 30° and 42°) and their response to different antibiotics.

Ribosomes from seventeen transductants with an altered response to temperature or antibiotics were isolated. They were tested for alterations in their ribosomal subunit profiles by sucrose centrifugation and for altered ribosomal proteins by twodimensional gel electrophoresis. Two strains showed accumulation of 50S ribosomal precursors and three strains had an altered 50S protein L18. This protein belongs to the 5S RNA-protein complex having GTPase and ATPase activity.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Buckel, P., Ruffler, D., Piepersberg, W., Böck, A.: RNA overproducing revertants of an analyl-tRNA synthetase mutant of Escherichia coli. Molec. gen. Genet. 119, 323 (1972)

    Google Scholar 

  • Davies, J., Nomura, M.: The genetics of bacterial ribosomes. Ann. Rev. Genet. 6, 203 (1972)

    Google Scholar 

  • Hong, J. S., Ames, B. N.: Localized mutagenesis of any specific small region of the bacterial chromosome. Proc. nat. Acad. Sci. (Wash.) 68, 3158 (1971)

    Google Scholar 

  • Horne, J. R., Erdmann, V. A.: ATPase and GTPase activities with a specific 5S RNA-protein complex. Proc. nat. Acad. Sci. (Wash.) 70, 2870 (1973)

    Google Scholar 

  • Jaskunas, S. R., Lindahl, L., Nomura, M.: Specialized transducing phages for ribosomal protein genes of Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 72, 6 (1975)

    Google Scholar 

  • Jaskunas, S. R., Nomura, M., Davies, J.: Genetics of bacterial ribosomes. In: Ribosomes, p. 333–368. Monograph Series: Cold Spring Harbor Laboratory 1974

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

    Google Scholar 

  • Lennox, E. X.: Transduction of linked genetic characters of the host by bacteriophage P1. Virology 1, 190 (1955)

    Google Scholar 

  • Miller, J. H.: Experiments in molecular genetics. p. 204: Cold Spring Harbor Laboratory 1972

  • Murgola, E., Yanofsky, C.: Structural interactions between amino acid residues at positions 22 and 211 in the tryptophan synthetase alpha chain of Escherichia coli. J. Bact. 117, 444 (1974)

    Google Scholar 

  • Osawa, S., Otaka, E., Takata, B., Dekio, S., Matsubara, M., Itoh, T., Muto, A., Tanaka, K., Teraoka, H., Tamaki, M.: Ribosomal protein genes in bacteria. FEBS Sympos. 23, 313 (1972)

    Google Scholar 

  • Stöffler, G.: Ribosomal proteins, II. A rapid micromethod for electrophoretic separation of E. coli ribosomal proteins on gelatinized cellulose acetate. Molec. gen. Genet. 100, 374 (1967)

    Google Scholar 

  • Stöffler, G., Wittmann, H. G.: Ribosomal proteins, XXV. Immunological studies on E. coli ribosomal proteins. J. molec. Biol. 62, 407 (1971a)

    Google Scholar 

  • Stöffler, G., Wittmann, H. G.: Ribosomal proteins, XXIII. Sequence differences of E. coli 30S ribosomal proteins as determined by immunochemical methods. Proc. nat. Acad. Sci. (Wash.) 68, 2283 (1971b)

    Google Scholar 

  • Strack, H. B., Bautz-Freese, E., Freese, E.: Comparison of mutations induced in bacteriophage and transforming DNA by various mutagens. Mutation Res. 1, 10 (1964)

    Google Scholar 

  • Wittmann, H. G., Stöffler, G., Geyl, D., Böck, A.: Alteration of ribosomal proteins in revertants of a valyl-tRNA synthetase mutant of Escherichia coli. Molec. gen. Genet., in press (1975)

  • Wittmann, H. G., Stöffler, G., Piepersberg, W., Buckel, P., Ruffler, D., Böck, A.: Altered S5 and S20 ribosomal proteins in revertants of an alanyl-tRNA synthetase mutant of Escherichia coli. Molec. gen. Genet. 134, 225 (1974)

    Google Scholar 

  • Wittmann, H. G., Wittmann-Liebold, B.: Chemical structure of bacterial ribosomal proteins. In: Ribosomes, p. 115–140. Monograph Series: Cold Spring Harbor Laboratory 1974

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by E. Bautz

Dedicated to Prof. G. Melchers on occasion of his 70th birthday.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Berger, I., Geyl, D., Böck, A. et al. Localized mutagenesis of the aroE-strA section of the Escherichia coli chromosome coding for ribosomal proteins. Molec. gen. Genet. 141, 207–211 (1975). https://doi.org/10.1007/BF00341800

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00341800

Keywords

Navigation