1887

Abstract

In K12 expression of the adenylate cyclase gene is subject to multiple controls. In order to gain understanding of the regulation of adenylate cyclase synthesis, operon and protein fusions were constructed by recombination either into bacteriophage or low-copy-number plasmids, or directly on the chromosome at the locus. The fusions were used in physiological experiments as probes to study transcriptional and translational controls of expression. It was found that adenylate cyclase synthesis was insensitive to glucose effects. As already described by other workers, the CAP-cAMP complex had a moderate negative control on expression. In addition it was observed that concomitant with a severe slackening of growth rate, specific to the growth of strains in rich medium, expression was considerably enhanced. This increase of adenylate cyclase synthesis did not appear to be directly dependent on the presence of a functional cAMP receptor (CAP), and seemed to be controlled at the level of transcription. Finally, translation of the message was very weak when compared to transcription (the mRNA level was the same in protein and operon fusions).

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-134-2-359
1988-02-01
2021-08-02
Loading full text...

Full text loading...

/deliver/fulltext/micro/134/2/mic-134-2-359.html?itemId=/content/journal/micro/10.1099/00221287-134-2-359&mimeType=html&fmt=ahah

References

  1. Aiba H. 1985; Transcription of the Escherichia coli adenylatecyclase gene is negatively regulated by cAMP-cAMP receptor protein. Journal of Biological Chemistry 5:3063–3070
    [Google Scholar]
  2. Aiba H., Kawamukai M., Ishihama A. 1983; Cloning and promoter analysis of the Escherichia coli adenylatecyclase gene. Nucleic Acids Research 11:3451–3465
    [Google Scholar]
  3. Aiba H., Mori K., Tanaka M., Ori T., Roy A., Danchin A. 1984; The complete nucleotide sequence of the adenylatecyclase gene of Escherichia coli. Nucleic Acids Research 12:9427–9439
    [Google Scholar]
  4. BirnboÎm H.C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research 7:1513–1523
    [Google Scholar]
  5. Botsford J.L. 1975; Metabolism of cyclic 3′-5′-monophosphate and induction of tryptophanase in Escherichia coli. Journal of Bacteriology 124:380–390
    [Google Scholar]
  6. Botsford J.L. 1981; Cyclic nucleotides in procaryotes. Microbiological Review 45:620–642
    [Google Scholar]
  7. Botsford J.L., Drexler M. 1978; The cyclic 3′- 5′-adenosine monophosphate receptor protein and regulation of cyclic 3′-5′-adenosine monophosphate synthesis in Escherichia coli. Molecular and General Genetics 165:47–65
    [Google Scholar]
  8. Coulondre C., Miller J.H. 1977; Genetic studies of the lac operon. III. Additional correlation of mutational sites with specific amino acid residues. Journal of Molecular Biology 117:525–575
    [Google Scholar]
  9. Danchin A., Guiso N., Roy A., Ullmann A. 1984; Identification of the Escherichia coli cya gene product as authentic adenylatecyclase. Journal of Molecular Biology 75:403–408
    [Google Scholar]
  10. Davis R.W., Botstein D. 1980 A Manual for Genetic Engineering: Advanced Bacterial Genetics. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  11. De Crombrugghe B., Busby S., Buc H. 1984; Cyclic AMP receptor protein: role in transcription activation. Science 224:831–837
    [Google Scholar]
  12. De Reuse H., Touati E., Glaser P., Danchin A. 1986; Low copy number plasmid vectors for gene cloning and for monitoring gene expression. FEMS Microbiology Letters 37:193–197
    [Google Scholar]
  13. Enquist L., Tiemer D., Leder P., Weisberg R., Steinberg N. 1976; Safer derivatives of bacteriophage lambda gt. Lambda C for use in cloning ofrecombinant DNA molecules. Nature; London: 259596–598
    [Google Scholar]
  14. Epstein W., Rothman-Denes L.B., Hesse J. 1975; cAMP as mediator of catabolite repression in Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America 72:2300–2304
    [Google Scholar]
  15. Gillepsie D., Spiegelman S. 1965; A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. Journal of Molecular Biology 12:829–842
    [Google Scholar]
  16. Guidi-Rontani C., Danchin A., Ullmann A. 1980; Catabolite repression in Escherichia coli mutants lacking cyclic AMP receptor protein. Proceedings of the National Academy of Sciences of the United States of America 77:5799–5801
    [Google Scholar]
  17. Guidi-Rontani C., Danchin A., Ullmann A. 1984; Transcriptional control of polarity in Escherichia coli by cAMP. Molecular and General Genetics 195:96–100
    [Google Scholar]
  18. Hagen F.S., Young E.T. 1978; Effect of RNAse III on efficiency of translation of bacteriophage T7 lysozyme mRNA. Journal of Virology 26:793–804
    [Google Scholar]
  19. Hohn B. 1979; In vitro packaging of λ and cosmid DNA. Methods in Enzymology 68:299–309
    [Google Scholar]
  20. Joseph E., Bernsley C., Guiso N., Ullmann A. 1982; Multiple regulation of the activity of adenylatecyclase in Escherichia coli. Molecular and General Genetics 185:262–268
    [Google Scholar]
  21. Kawamukai M., Kishimoto J., Utsumi R., Himeno M., Komano T., Aiba H. 1985; Negative regulation of adenylatecyclase gene (cya) expression by cyclic AMP-cyclic AMP receptor protein in Escherichia coli: studies with cya-lac protein and operon fusion plasmids. Journal of Bacteriology 164:872–877
    [Google Scholar]
  22. Kourilsky P., Mercereau O., Gros D., Tremblay G. 1974; Hybridization on filters with competitor DNA in the liquid phase in a standard and a micro-assay. Biochimie 56:1215–1221
    [Google Scholar]
  23. Mackie G.A. 1981; Nucleotide sequence of the gene for ribosomal protein S20 and its flanking regions. Journal of Biological Chemistry 256:8177–8182
    [Google Scholar]
  24. Matin A., Matin M.K. 1982; Cellular levels, excretion and synthesis rates of cyclic AMP in Escherichia coli grown in continuous culture. Journal of Bacteriology 149:801–807
    [Google Scholar]
  25. Miller J.H. 1972 Experiments in Molecular Genetics. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  26. Pardee A.B., Jacob F., Monod J. 1959; The genetic control and cytoplasmic expression of induc- ibility in the synthesis of beta-galactosidase of Escherichia coli. Journal of Molecular Biology 1:165–168
    [Google Scholar]
  27. Potter K., Chalmers-Larson G., Yamazaki H. 1974; Abnormally high rate of cyclic AMP excretion from an Escherichia coli mutant deficient in cyclic AMP receptor protein. Biochemical and Biophysical Research Communications 57:379–386
    [Google Scholar]
  28. Reddy P., Peterkofsky A., Mckenney K. 1985; Translational efficiency of the Escherichia coli adenylatecyclase gene: mutating the UUG initiation codon to GUG or AUG results in increased gene expression. Proceedings of the National Academy of Sciences of the United States of America 82:5656–5660
    [Google Scholar]
  29. Roy A., Danchin A. 1982; The cya locus of Escherichia coli K12: organization and gene products. Molecular and General Genetics 188:465–471
    [Google Scholar]
  30. Roy A., Danchin A., Joseph E., Ullmann A. 1983a; Two functional domains in adenylatecyclase of Escherichia coli. Journal of Molecular Biology 165:197–202
    [Google Scholar]
  31. Roy A., Haziza C., Danchin A. 1983b; Regulation of adenylatecyclase synthesis in Escherichia coli: nucleotide sequence of the control region. EMBO Journal 2:791–797
    [Google Scholar]
  32. Stormo G.D., Schneider T.D., Gold L., Ehrenfeucht A. 1982; Use of the ‘Perceptron’ algorithm to distinguish translational initiation sites in E.coli. Nucleic Acids Research 10:2997–3011
    [Google Scholar]
  33. Ullmann A. 1984; One-step purification of hybrid proteins which have beta-galactosidase activity. Gene 29:27–31
    [Google Scholar]
  34. Ullmann A., Danchin A. 1983; Role of cyclic AMP in bacteria. Advances in Cyclic Nucleotides Research 15:1–53
    [Google Scholar]
  35. Ullmann A., Tillier F., Monod J. 1976; Catabolite modulator factor: a possible mediator of catabolite repression in bacteria. Proceedings of the National Academy of Sciences of the United States of America 73:3476–3479
    [Google Scholar]
  36. Ullmann A., Joseph E., Danchin A. 1979; Cyclic AMP as a modulator of polarity in polycis- tronic transcriptional units. Proceedings of the National Academy of Sciences of the United States of America 76:3194–3197
    [Google Scholar]
  37. Wang J.Y.J., Clegg D.O., Koshland D.E.Jr 1981; Molecular cloning and amplification of the adenylatecyclase gene. Proceedings of the National Academy of Sciences of the United States of America 78:4684–4688
    [Google Scholar]
  38. Wayne P.K., Rosen O.M. 1974; Cyclic 3′-5′-adenosine monophosphate in Escherichia coli during transient and catabolite repression. Proceedings of the National Academy of Sciences of the United States of America 71:1436–1440
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-134-2-359
Loading
/content/journal/micro/10.1099/00221287-134-2-359
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error