Skip to main content
Log in

Involvement of host factors in the regulation of the Vibrio fischeri lux operon in Escherichia coli cells

  • Conference Proceedings
  • Published:
Microbiology Aims and scope Submit manuscript

Abstract

It has been shown that the chaperonin GroEL, together with GroES co-chaperonin and Lon ATP-dependent protease are involved in the regulation of expression of the Vibrio fischeri lux operon in Escherichia coli cells. The cells of E. coli groE (pF1)−1 bearing a plasmid with the complete V. fischeri lux regulon were weakly luminescent. The cells of E. coli lonA (pF1) displayed intense bioluminescence. The same effects also occurred in mutant E. coli strains bearing a hybrid plasmid pVFR1, where the luxR gene and the regulatory region of the V. fischeri lux operon were inserted before the Photorhabdus luminescens luxCDABE cassette. The V. fischeri luxR gene was cloned in the pGEX-KG vector with the formation of a hybrid gene gst-luxR. It was shown that affinity chromatography of the product of expression, the chimeric protein GST-LuxR, on a column with glutathione-agarose resulted in its copurification with the proteins GroEL and Lon. Consequently, LuxR, the transcription activator of the lux operon, forms complexes with these proteins. It is supposed that GroEL/GroES is responsible for the folding of the LuxR protein, and Lon protease degrades the LuxR protein either before its folding into an active globule or at denaturing.

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.

Institutional subscriptions

Similar content being viewed by others

References

  1. Fuqua, W.C., Winans, S.C., and Greenberg, E.P., Quorum Sensing in Bacteria: the LuxR-LuxI Family of Cell Density-Responsive Transcriptional Regulators, J. Bacteriol., 1994, vol. 176, pp. 269–275.

    PubMed  CAS  Google Scholar 

  2. Ulitzur, S. and Dunlop, P., Regulatory Circuitry Controlling Luminescence Autoinduction in Vibrio fischeri, Photochem. Photobiol., 1995, vol. 62, pp. 625–632.

    CAS  Google Scholar 

  3. Urbanowski, M.L., Lostroh, C.P., and Greenberg, E.P., Reversible Acyl-Homoserine Lactone Binding to Purified Vibrio fischeri LuxR Protein, J. Bacteriol., 2004, vol. 186, pp. 631–637.

    Article  PubMed  CAS  Google Scholar 

  4. Dunlop, P.V. and Greenberg, E.P., Control of Vibrio fischeri lux Genes Transcription by CAMP-Receptor Protein-LuxR Regulatory Circuit, J. Bacteriol., 1988, vol. 170, pp. 4040–4046.

    Google Scholar 

  5. Ulitzur, S., Matin, A., Fraley, C., and Meighen, E.A., H-NS Protein Represses Transcription of the lux-Systems of Vibrio fischeri and Others Luminous Bacteria Cloned into Escherichia coli, Curr. Microbiol., 1997, vol. 35, pp. 336–342.

    Article  PubMed  CAS  Google Scholar 

  6. Zavil’gel’skii, G.B. and Kotova, V.Yu., Osmotic Shock Induction of the Expression of Vibrio fischeri lux Genes in Escherichia coli Cells, Genetika, 2003, vol. 39, no. 4, pp. 483–488 [Russ. J. Genet. (Engl. Transl.), vol. 39, no. 4, pp. 390–394].

    PubMed  CAS  Google Scholar 

  7. Dolan, K.M. and Greenberg, E.P., Evidence That GroEL, notσ32, Is Involved in Transcriptional Regulation of the Vibrio fischeri Luminescence Genes in Escherichia coli, J. Bacteriol., 1992, vol. 174, pp. 5132–5135.

    PubMed  CAS  Google Scholar 

  8. Adar, Y.Y., Simaan, M., and Ulitzur, S., Formation of the LuxR Protein in the Vibrio fischeri lux System Is Controlled by HtprR Thorough the GroESL Proteins, J. Bacteriol., 1992, vol. 174, pp. 7138–7143.

    PubMed  CAS  Google Scholar 

  9. Zavil’gel’skii, G.B. and Manukhov, I.V., Lon Protease Participates in the Regulation of lux Operon Transcription in Vibrio fischeri, Genetika, 1994, vol. 30, pp. 337–341.

    PubMed  CAS  Google Scholar 

  10. Ulitzur, S., Involvement of Host Factors in Regulating the lux System of Vibrio fischeri., Microbial Ecology and Infectious Disease, Rosenberg, E., Ed., Washington: Am. Soc. Microbiol, 1999, pp. 123–132.

    Google Scholar 

  11. Zeilstra-Ryalls, J., Fayet, O., Baird, L., and Georgopoulos, C., Sequence Analysis and Phenotypic Characterization of groEL Mutations that Block λ and T4 Bacteriophage Growth, J. Bacteriol., 1993, vol. 175, no. 4, pp. 1134–1143.

    PubMed  CAS  Google Scholar 

  12. Guan, K.L. and Dixon, J.E., Eukaryotic Proteins Expressed in Escherichia coli: an Inproved Thrombin Cleavage and Purification Procedure of Fusion Proteins with Glutathione S-Transferase, Anal. Biochem., 1991, vol. 192, pp. 262–267.

    Article  PubMed  CAS  Google Scholar 

  13. Van Dyk, T.K. and Rosson, R.A., Photorhabdus luminescens luxCDABE Promoter Probe Vectors, Methods in Molecular Biology, Larossa, R.A., Ed., Humana Press Inc, 1998, vol. 102, pp. 85–95.

  14. Rosenfeld, J., Capdevielle, J., Guillemoi, J., and Ferrara, P., In-Gel Digestion of Proteins for Internal Sequence Analysis After One-Or Two Dimensional Gel Electrophoresis, Anal. Biochem., 1992, vol. 203, pp. 173–179.

    Article  PubMed  CAS  Google Scholar 

  15. Patterson, S.D. and Aebersold, R., Mass-Spectrometric Approaches for the Identification of Gel-Separated Proteins, Electrophoresis, 1995, vol. 16, pp. 1791–1814.

    Article  PubMed  CAS  Google Scholar 

  16. Bairoch, A. and Apweiler, R., The SWISS-PROT Protein Sequence Database and Its Supplement TrEMBL 2000, Nucleic Acid Res., 2000, vol. 28, pp. 45–48.

    Article  PubMed  CAS  Google Scholar 

  17. Baneyx, F. and Gatenby, A.A., A Mutation in GroEL Interferes with Protein Folding by Reduction the Rate of Discharge of Sequestered Polypeptides, J. Biol. Chem., 1992, vol. 267, no. 16, pp. 11637–11644.

    PubMed  CAS  Google Scholar 

  18. Zhu, J. and Winans, S.C., The Quorum-Sensing Transcriptional Regulator TraR Requires Its Cognate Signaling Ligand for Protein Folding, Protease Resistance, and Dimerization, Proc. Natl. Acad. Sci. USA, 2001, vol. 98, pp. 1507–1512.

    Article  PubMed  CAS  Google Scholar 

  19. Zhu, J., Oger, P.M., Schrammeijer, B., Hooykaas, P.J.J., Farrand, S.K., and Winans, S.C., The Bases of Crown Gall Tumorigenesis, J. Bacteriol., 2000, vol. 182, pp. 3885–3895.

    Article  PubMed  CAS  Google Scholar 

  20. Ruby, E.G., Urbanowski, M., Campbell, J., Dunn, J.A., Faini, M., Gunsalus, R., Lostroh, P., Lupp, C., McCann, J., Millikan, D., Schaefer, A., Stabb, E., Stevens, A., Visick, K., Whistler, C., and Greenberg, E.P., Complete Genome Sequence of Vibrio fischeri: a Symbiotic Bacterium with Pathogenic Congeners, Proc. Natl. Acad. Sci. USA, 2005, vol. 102, no. 8, pp. 3004–3009.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to G. B. Zavil’gel’sky.

Additional information

Original Russian Text © I.V. Manukhov, V.Yu. Kotova, G.B. Zavil’gel’sky, 2006, published in Mikrobiologiya, 2006, Vol. 75, No. 4, pp. 525–531.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Manukhov, I.V., Kotova, V.Y. & Zavil’gel’sky, G.B. Involvement of host factors in the regulation of the Vibrio fischeri lux operon in Escherichia coli cells. Microbiology 75, 452–458 (2006). https://doi.org/10.1134/S002626170604014X

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S002626170604014X

Key words

Navigation