Skip to main content
SpringerLink
Log in

A transcriptional barrier in the regulatory region of the tryptophan operon of Escherichia coli: Its role in the regulation of repressor-independent RNA synthesis

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

Summary

A study was made of the influence of the growth rate of Escherichia coli bacteria on the transcription of the tryptophan (trp) operon, in various trp repressor negative strains. Selective measurement of the levels of RNA transcribed from the regulatory region (reg) of this operon and from the structural genes, respectively, has revealed that the increase of the rate of trpRNA1 synthesis with bacterial growth rate (Rose and Yanofsky, 1972) is due to a decrease of the frequency of termination of transcription at the transcriptional barrier in the regulatory region of the operon.

In a mutant strain of E. coli with a deletion covering the promotor distal part of the regulatory region of the trp operon where the barrier is located, the rate of trpRNA synthesis is not affected by the growth rate.

In suA − strains, in which Rho factor activity is reduced the bacterial growth rate does not affect the rate of synthesis of trpRNA. This result suggests that in wild type bacteria Rho factor contributes to the control of the transcription of the trp operon. In bacteria with a mutation rendering Tryptophanyl-tRNA synthetase (TRSase) inactive (trpS − strains) the rate of trpRNA synthesis is affected by the growth rate in the same way as in the isogenic wild type bacteria. This result indicates that TRSase plays no obligatory role in the control of trpRNA synthesis through a mechanism of termination and anti-termination of transcription, at least not in the studied strains, which carried a relA mutation.

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

  • Adhya, S., Gottesman, M., Crombrugghe, B. de: Release of polarity in Escherichia coli by gene N of phage λ: Termination and anti-termination of transcription. Proc. nat. Acad. Sci. (Wash.) 71, 2534–2538 (1974)

    Google Scholar 

  • Bertrand, K., Korn, L., Lee, F., Platt, T., Squires, C.L., Squires, C., Yanofsky, C.: New features of the regulation of the tryptophan operon. Science 189, 22–26 (1975)

    Google Scholar 

  • Bertrand, K., Squires, C., Yanofsky, C.: Transcription termination in vivo in the leader region of the tryptophan operon of Escherichia coli. J. molec. Biol. 103, 319–337 (1976)

    Google Scholar 

  • Bertrand, K., Yanofsky, C.: Regulation of transcription termination in the leader region of the tryptophan operon of Escherichia coli involves tryptophan or its metabolic product. J. molec. Biol. 103, 339–349 (1976)

    Google Scholar 

  • Camakaris, J., Pittard, J.: Repression of 3-Deoxy-D-Arabinoheptulosine Acid-7-Phosphate Synthetase (trp) and enzymes of the tryptophan pathway in Escherichia coli K-12. J. Bact. 107, 406–414 (1971)

    Google Scholar 

  • Cohen, G., Jacob, F.: Sur la répression de la synthèse des enzymes intervenant dans la formation du tryptophane chez Escherichia coli. C.R. Acad. Sci. (Paris) 248, 3490–3492 (1959)

    Google Scholar 

  • Crombrugge, B. de, Adhya, S., Gottesman, M., Pastan, I.: Effect of Rho on transcription of bacterial operons. Nature (Lond.) New Biol. 241, 260–264 (1973)

    Google Scholar 

  • Davison, J., Brammar, W.J., Brunel, F.: Quantitative aspects of gene expression in a λ-trp fusion operon. Molec. gen. Genet. 130, 9–20 (1974)

    Google Scholar 

  • Deeb, S.S., Okamoto, K., Hall, B.: Isolation and characterization of non-defective transducing elements of bacteriophage ϕ80. Virology 31, 289–295 (1967)

    Google Scholar 

  • Doolittle, W.F., Yanofsky, C.: Mutants of Escherichia coli with an altered Tryptophanyl-Transfer Ribonucleic Acid Synthetase. J. Bact. 95, 1283–1294 (1968)

    Google Scholar 

  • Franklin, N.C.: Altered reading of genetic signals fused to the N operon of bacteriophage λ: Genetic evidence for modification of Polymerase by the protein product of the N gene. J. molec. Biol. 89, 33–48 (1974)

    Google Scholar 

  • Franklin, N.C., Dove, W.F., Yanofsky, C.: A linear insertion of a prophage into the chromosome of E. coli shown by deletion mapping. Biochem. biophys. Res. Commun. 18, 1910–1923 (1965)

    Google Scholar 

  • Gillespie, D., Spiegelman, S.: A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J. molec. Biol. 12, 829–842 (1965)

    Google Scholar 

  • Goff, C.G., Minkley, E.G.: The RNA polymerase sigma factor: a specificity determinant. Lepetit colloquia on biology and medicine vol. I (L. Silvestri ed.), pp. 124–147. Amsterdam: North-Holland 1970

    Google Scholar 

  • Ito, K.: Regulatory mechanism of the tryptophan operon in Escherichia coli: Possible interaction between trpR and trpS gene products. Molec. gen. Genet. 115, 349–363 (1972)

    Google Scholar 

  • Ito, K., Hiraga, S., Yura, T.: Temperature-sensitive repression of the tryptophan operon in Escherichia coli. J. Bact. 99, 279–286 (1969a)

    Google Scholar 

  • Ito, K., Hiraga, S., Yura, T.: Tryptophanyl transfer RNA synthetase and expression of the tryptophan operon in the trpS mutants of Escherichia coli. Genetics 61, 521–538 (1969b)

    Google Scholar 

  • Jackson, E.N., Yanofsky, C.: The region between the operator and first structural gene of the tryptophan operon of Escherichia coli may have a regulatory function. J. molec. Biol. 76, 89–101 (1973)

    Google Scholar 

  • Kano, Y., Matsushiro, A., Shimura, Y.: Isolation of the novel regulatory mutants of the tryptophan biosynthetic system in Escherichia coli. Molec. gen. Genet. 102, 15–26 (1968)

    Google Scholar 

  • Kuwano, M., Schlessinger, D., Morse, D.: Loss of dispensable endonuclease activity in relief of polarity by suA. Nature (Lond.) New Biol. 231, 214–217 (1971)

    Google Scholar 

  • Lee, F., Squires, C.L., Squires, C., Yanofsky, C.: Termination of transcription in vitro in the Escherichia coli tryptophan operon leader region. J. molec. Biol. 103, 383–393 (1976)

    Google Scholar 

  • Maaløe, O., Kjeldgaard, N.O.: Control of macromolecular synthesis. New York: W.A. Benjamin, Inc. 1966

    Google Scholar 

  • McGeoch, D., McGeoch, J., Morse, D.: Synthesis of Tryptophan operon RNA in a cell-free system. Nature (Lond.) New Biol. 245, 137–140 (1973)

    Google Scholar 

  • Morse, D., Morse, A.: Dual control of the tryptophan operon by the Repressor and Tryptophanyl-tRNA Synthetase. J. molec. Biol. 103, 209–226 (1976)

    Google Scholar 

  • Mosteller, R.D., Rose, J.K., Yanofsky, C.: Transcription initiation and degradation of trp mRNA. Cold Spr. Harb. Symp. quant. Biol. 35, 461–466 (1970)

    Google Scholar 

  • Murray, N.E., Brammar, W.J.: The trpE gene of Escherichia coli K contains a recognition sequence for the K-restriction system. J. molec. Biol. 77, 615–624 (1973)

    Google Scholar 

  • Pannekoek, H., Brammar, W.J., Pouwels, P.H.: Punctuation of transcription in vitro of the tryptophan operon of Escherichia coli. A novel type of control of transcription. Molec. gen. Genet. 136, 199–214 (1975)

    Google Scholar 

  • Pannekoek, H., Perbal, B., Pouwels, P.: The specificity of transcription in vitro of the Tryptophan operon of Escherichia coli. II. The effect of Rho factor. Molec. gen. Genet. 132, 291–306 (1974)

    Google Scholar 

  • Pannekoek, H., Pouwels, P.H.: The specificity of transcription in vitro of the trp operon of Escherichia coli tryptophan operon messenger RNA. J. molec. Biol. 103, 411–420 (1976)

    Google Scholar 

  • Pouwels, P.H., Rotterdam, J. van: In vitro synthesis of enzymes of the tryptophan operon of Escherichia coli. III. Evidence for positive control of transcription. Molec. gen. Genet. 136, 185–197 (1975)

    Google Scholar 

  • Ratner, D.: Evidence that mutations in the suA polarity suppressing gene directly affect termination factor Rho. Nature (Lond.) 259, 151–153 (1976)

    Google Scholar 

  • Richardson, J.P., Grimley, C., Lowery, C.: Transcription termination factor Rho activity is altered in Escherichia coli with suA gene mutations. Proc. nat. Acad. Sci. (Wash.) 72, 1725–1728 (1975)

    Google Scholar 

  • Roberts, J.W.: Termination factor for RNA synthesis. Nature (Lond.) 224, 1168–1171 (1969)

    Google Scholar 

  • Roberts, J.W.: Transcription termination and late control in phage lambda. Proc. nat. Acad. Sci. (Wash.) 72, 3300–3304 (1975)

    Google Scholar 

  • Rose, J.K., Mosteller, R.D., Yanofsky, C.: Tryptophan messenger ribonucleic acid elongation rates and steady-state levels of tryptophan operon enzymes under various growth conditions. J. molec. Biol. 51, 541–550 (1970)

    Google Scholar 

  • Rose, J.K., Squires, C.L., Yanofsky, C., Yang, H.-L., Zubay, G.: Regulation of in vitro transcription of the Tryptophan operon by purified RNA polymerase in the presence of partially purified repressor and Tryptophan. Nature (Lond.) New Biol. 245, 133–137 (1973)

    Google Scholar 

  • Rose, J.K., Yanofsky, C.: Metabolic regulation of the Tryptophan operon of Escherichia coli: Repressor-independent regulation of transcription initiation frequency. J. molec. Biol. 69, 103–118 (1972)

    Google Scholar 

  • Rosenberg, M., Weissman, S., Crombrugghe, B. de: Termination of transcription in bacteriophage lambda. Heterogeneous 3′-terminal oligoadenylate additions and the effect of ρ factor. J. biol. Chem 250, 4755–4764 (1975)

    Google Scholar 

  • Segawa, T., Imamoto, F.: Diversity of regulation of genetic transcription. II. Specific relaxation of polarity in read-through transcription of the translocated trp operon in bacteriophage lambda trp. J. molec. Biol. 87, 741–734 (1974)

    Google Scholar 

  • Squires, C.L., Rose, J.K., Yanofsky, C., Yang, H.L., Zubay, G.: Tryptophanyl-tRNA and Tryptophanyl-tRNA synthetase are not required for in vitro repression of the tryptophan operon. Nature (Lond.) New Biol. 245, 131–133 (1973)

    Google Scholar 

  • Summers, W.C.: A simple method for extraction of RNA from E. coli utilizing diethyl pyrocarbonate.

  • Vogel, H., Bonner, D.M.: Acetylornithinase of Escherichia coli: partial purification and some properties. J. biol. Chem. 218, 97–106 (1956)

    Google Scholar 

Download references

Author information

Author notes

  1. Hans Pannekoek

    Present address: Laboratory for Molecular Genetics, State University of Lieden, The Netherlands

Authors and Affiliations

  1. Medical Biological Laboratory TNO, 2100, Rijswijk, The Netherlands

    Peter H. Pouwels & Hans Pannekoek

Authors
  1. Peter H. Pouwels
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hans Pannekoek
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by E. Bautz

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pouwels, P.H., Pannekoek, H. A transcriptional barrier in the regulatory region of the tryptophan operon of Escherichia coli: Its role in the regulation of repressor-independent RNA synthesis. Molec. Gen. Genet. 149, 255–265 (1976). https://doi.org/10.1007/BF00268526

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00268526

Keywords

Search

Navigation