Skip to main content
SpringerLink
Log in

Deductive analysis of a protein-synthesis mutant of Escherichia coli

  • Published:
Biochemical Genetics Aims and scope Submit manuscript

Abstract

A mutant of E. coli, T s68b, selected as unable to grow at 43 C, is unable to synthesize proteins at 43 C, though it can carry out this function at 30 C. This defect is shown to be in the protein-synthetic rather than the RNA-synthetic machinery by an analysis of the response of the strain to infection with the RNA bacteriophage f2. An analysis of the capacity for RNA synthesis and the polyribosome content of these cells at 44 C indicates that the defect resides in the elongation step of protein synthesis. No defects could be detected in vitro. The results are discussed in light of similar data on other mutants and in relation to the general approach of analyzing complex mutants selected with ill-defined phenotypes.

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

  • Adelberg, E., Mandel, M., and Chen, G. C. (1965). Optimal conditions for mutagenesis by N-methyl′-nitro-N-nitrosoguanidine in Escherichia coli K12. Biochem. Biophys. Res. Commun. 18788.

    Google Scholar 

  • Apirion, D. (1967). Three genes that affect Escherichia coli ribosomes. J. Mol. Biol. 30255.

    Google Scholar 

  • Austin, S. J., Tittawalla, I. R., Hayward, R. S., and Scaife, J. (1971). Amber mutations of Escherichia coli RNA polymerase. Nature New Biol. 232133.

    Google Scholar 

  • Beller, R. J., and Davis, B. D. (1971). Selective dissociation of free ribosomes of Escherichia coli by sodium ions. J. Mol. Biol. 55477.

    Google Scholar 

  • Bowman, C. M., Dahlberg, J. E., Ikemura, T., Konisky, J., and Nomura, M. (1971). Specific inactivation of 16S ribosomal RNA induced by colicin E3 in vivo. Proc. Natl. Acad. Sci. 68964.

    Google Scholar 

  • Boyd, D., Nixon, R., Gillespie, S., and Gillespie, D. (1968). Screening of Escherichia coli temperature-sensitive mutants by pretreatment with glucose starvation. J. Bacteriol. 951040.

    Google Scholar 

  • Cohen, L. B., Goldberg, J. H., and Herner, A. E. (1969). Inhibition by pactamycin of the initiation of protein synthesis. Effect on the 30S ribosomal subunit. Biochemistry 81312.

    Google Scholar 

  • Eidlic, L., and Neidhardt, F. C. (1965). Protein and nucleic acid synthesis in two mutants of Escherichia coli with temperature-sensitive amino acyl ribonucleic acid synthetases. J. Bacteriol. 89706.

    Google Scholar 

  • Fedoroff, N., and Zinder, N. D. (1971). Structure of the poly (G) polymerase component of the bacteriophage f2 replicase. Proc. Natl. Acad. Sci. 681838.

    Google Scholar 

  • Friesen, J. D. (1969). Dependence of f2 bacteriophage RNA replication on amino acids. J. Mol. Biol. 46349.

    Google Scholar 

  • Fromageot, H. P. M., and Zinder, N. D. (1968). Growth of bacteriophage f2 in E. coli treated with rifampicin. Proc. Natl. Acad. Sci. 61184.

    Google Scholar 

  • Godson, G. N., and Sinsheimer, R. L. (1967). Lysis of E. coli with a neutral detergent. Biochim. Biophys. Acta 149476.

    Google Scholar 

  • Gurgo, C., Apirion, D., and Schlessinger, D. (1969a). Effects of chloramphenicol and fusidic acid on polyribosome metabolism in Escherichia coli. FEBS Letters 334.

    Google Scholar 

  • Gurgo, C., Apirion, D., and Schlessinger, D. (1969b). Polyribosome metabolism in E. coli treated with chloramphenicol, neomycin, spectinomycin, or retracycline. J. Mol. Biol. 45205.

    Google Scholar 

  • Gurgo, C., Craig, E., Schlessinger, D., and Afolayan, A. (1972). Polyribosome metabolism in Escherichia coli starved for an amino acid. J. Mol. Biol. 62525.

    Google Scholar 

  • Hong, J. S., and Ames, B. N. (1971). Localized mutagenesis of any specific small region of the bacterial chromosome. Proc. Natl. Acad. Sci. 683158.

    Google Scholar 

  • Hori, M., Kunimoto, T., and Suzuki, J. (1969). Rapidly reversible dissociation of polysomes in Escherichia coli by n-butanol. J. Biochem. 66705.

    Google Scholar 

  • Jacobson, A. (1971). Studies of a temperature-sensitive RNA polymerase mutant of Escherichia coli. Ph.D. thesis, Brandeis University.

  • Jacobson, A., and Gillespie, D. (1968). Metabolic events occurring during recovery from prolonged glucose starvation in Escherichia coli. J. Bacteriol. 951030.

    Google Scholar 

  • Jacobson, A., and Gillespie, D. (1970). An RNA polymerase mutant defective in ATP initiations. Cold Spring Harbor Symp. Quant. Biol. 3585.

    Google Scholar 

  • Jacobson, A., and Gillespie, D. (1971). Genetic complementation between Escherichia coli RNA polymerase mutants. Biochem. Biophys. Res. Commun. 441030.

    Google Scholar 

  • Kaempfer, R. O. R. (1970). Dissociation of ribosomes on polypeptide chain termination and the origin of single ribosomes. Nature 228534.

    Google Scholar 

  • Kamen, R. (1970). Characterization of the subunits of QB replicase. Nature 228527.

    Google Scholar 

  • Kondo, M., Gallerani, R., and Weissman, C. (1970). Subunit structure of the QB replicase. Nature 228525.

    Google Scholar 

  • Lodish, H. F. (1970). Secondary structure of bacteriophage f2 ribonucleic acid and the initiation of in vitro protein biosynthesis. J. Mol. Biol. 50689.

    Google Scholar 

  • Lodish, H. F., Housman, D., and Jacobsen, M. (1971). Initiation of hemoglobin synthesis. Specific inhibition by antibiotics and bacteriophage RNA. Biochemistry 102348.

    Google Scholar 

  • Lowry, O. H., Rosebrough, N., Farr, A., and Randall, R. (1951). Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193265.

    Google Scholar 

  • Maruta, H., Natori, S., and Mizuno, D. J. (1969). Protein synthesis with E. coli ribosomes altered in conformation by monovalent cations. J. Mol. Biol. 46513.

    Google Scholar 

  • Modellel, J., and Davis, B. D. (1968). Rapid inhibition of polypeptide chain extension by streptomycin. Proc. Natl. Acad. Sci. 611279.

    Google Scholar 

  • Modollel, J., and Davis, B. D. (1969). A unitary mechanism for the several effects of streptomycin on the ribosome. Cold Spring Harbor Symp. Quant. Biol. 34113.

    Google Scholar 

  • Neidhardt, F. C. (1966). Roles of amino acid activating enzymes in cellular physiology. Bacteriol. Rev. 30701.

    Google Scholar 

  • Nierlich, D. P. (1967). Radioisotope uptake as a measure of synthesis of messenger RNA. Science 1581186.

    Google Scholar 

  • Noll, H., Staehelin, T., and Wettstein, F. O. (1963). Ribosomal aggregates engaged in protein synthesis: Ergosome breakdown and messenger ribonucleic acid transport. Nature 198632.

    Google Scholar 

  • Patterson, D. (1971). Studies on protein synthesis in temperature-sensitive mutants of E. coli. Ph.D. thesis, Brandeis University.

  • Patterson, D., and Gillespie, D. (1971). Stringent response of RNA synthesis in E. coli. produced by a temperature shift-up. Biochem. Biophys. Res. Commun. 45476.

    Google Scholar 

  • Patterson, D., and Gillespie, D. (1972). The effect of elevated temperatures on protein synthesis in E. coli. J. Bacteriol. 112 (in press).

  • Patterson, D., Weinstein, M., Marshall, S., and Gillespie, D. (1971). A new RNA synthesis mutant of E. coli. Biochem. Genet. 5563.

    Google Scholar 

  • Revel, M., Herzberg, M., Becarevic, A., and Gros, F. (1968). Role of a protein factor in the functional binding of ribosomes to natural messenger RNA. J. Mol. Biol. 33231.

    Google Scholar 

  • Rose, J., Mosteller, R., and Yanofsky, C. (1970). Tryptophan messenger ribonucleic acid elongation rates and steady-state levels of tryptophan operon enzymes under various growth conditions. J. Mol. Biol. 51541.

    Google Scholar 

  • Scheps, R., Wax, R., and Revel, M. (1971). Reactivation in vitro of inactive ribosomes from stationary phase Escherichia coli. Biochim. Biophys. Acta 232140.

    Google Scholar 

  • Senior, B. W., Kwasniak, J., and Holland, J. B. (1970). Colicin E3-directed changes in ribosome function and polyribosome metabolism in Escherichia coli K12. J. Mol. Biol. 53205.

    Google Scholar 

  • Stent, G. S., and Brenner, S. (1961). A genetic locus for the regulation of ribonucleic acid synthesis. Proc. Natl. Acad. Sci. 472005.

    Google Scholar 

  • Stewart, M. L., Grollman, A. P., and Huang, M. T. (1971). Aurintricarboxylic acid: Inhibitor of initiation of protein synthesis. Proc. Natl. Acad. Sci. 6897.

    Google Scholar 

  • Taylor, A. L., and Trotter, C. D. (1967). Revised linkage map of Escherichia coli. Bacteriol. Rev. 31332.

    Google Scholar 

  • Teraoka, H. (1970). A reversible change in the ability of E. coli ribosomes to bind to erythromycin. J. Mol. Biol. 48511.

    Google Scholar 

  • Tocchini-Valentini, G., and Mattocia, E. (1968). A mutant of E. coli with an altered supernatant factor. Proc. Natl. Acad. Sci. 61146.

    Google Scholar 

  • Vassart, G. M., Dumont, J. E., and Cantraine, F. R. L. (1970). Simulation of polyribosome disaggregation. Biochim. Biophys. Acta 224155.

    Google Scholar 

  • Weber, K., and Osborn, M. (1969). The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J. Biol. Chem. 2444406.

    Google Scholar 

  • Webster, R. E., and Zinder, N. D. (1969). Fate of the message-ribosome complex upon translation of termination signals. J. Mol. Biol. 42425.

    Google Scholar 

  • Weisblum, B., and Davies, J. (1968). Antibiotic inhibitors of the bacterial ribosome. Bacteriol. Rev. 32493.

    Google Scholar 

  • Young, R. M., and Nakada, D. (1971). Defective ribosomes in chloramphenicol-treated E. coli. J. Mol. Biol. 57457.

    Google Scholar 

Download references

Author information

Author notes

  1. David Patterson

    Present address: Department of Biophysics, University of Colorado Medical Center, Denver, Colorado

  2. David Gillespie

    Present address: Gerontology Research Center, Baltimore City Hospitals, Baltimore, Maryland

Authors and Affiliations

  1. Biology Department, Brandeis University, Waltham, Massachusetts, and Laboratory of Molecular Aging, Gerontology Research Center, NICHD, NIH, and the Baltimore City Hospitals, Baltimore, Maryland

    David Patterson & David Gillespie

Authors
  1. David Patterson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. David Gillespie
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This work was supported by Grant GM14368 from the National Institute of Health.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Patterson, D., Gillespie, D. Deductive analysis of a protein-synthesis mutant of Escherichia coli . Biochem Genet 8, 205–230 (1973). https://doi.org/10.1007/BF00485547

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00485547

Keywords

Search

Navigation