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. 1997 Jan;179(2):557–562. doi: 10.1128/jb.179.2.557-562.1997

Quorum sensing in Vibrio fischeri: essential elements for activation of the luminescence genes.

A M Stevens 1, E P Greenberg 1
PMCID: PMC178731  PMID: 8990313

Abstract

LuxR is required for cell density-dependent activation of the Vibrio fischeri luminescence (lux) genes. It has not been possible to study full-length LuxR in vitro, but a polypeptide containing the C-terminal transcriptional-activator domain of LuxR (LuxRdeltaN) has been purified, and its binding to lux regulatory DNA has been investigated. By itself, LuxRdeltaN interacts with a region of lux regulatory DNA that is upstream of the lux box, which is a 20-bp element that is required for LuxR activation of the luminescence operon. Individually, neither the purified LuxRdeltaN nor RNA polymerase binds to the lux box region, but together the two proteins bind in synergy to the lux box-luxI promoter region. We show that binding of LuxRdeltaN to the upstream region is not a prerequisite for its synergistic binding with RNA polymerase to the lux box and the luxI promoter region. We also show that LuxRdeltaN and RNA polymerase are both required and sufficient for transcriptional activation of the lux operon. This argues against the hypothesis that LuxR functions to alleviate repression of the lux operon by another cellular factor. Rather, our data support the view that LuxR functions as an accessory factor that enables RNA polymerase to bind to and initiate transcription from the promoter of the lux operon.

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Selected References

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  1. Brennan R. G., Matthews B. W. The helix-turn-helix DNA binding motif. J Biol Chem. 1989 Feb 5;264(4):1903–1906. [PubMed] [Google Scholar]
  2. Chen Y. H., Yang J. T., Chau K. H. Determination of the helix and beta form of proteins in aqueous solution by circular dichroism. Biochemistry. 1974 Jul 30;13(16):3350–3359. doi: 10.1021/bi00713a027. [DOI] [PubMed] [Google Scholar]
  3. Choi S. H., Greenberg E. P. Genetic dissection of DNA binding and luminescence gene activation by the Vibrio fischeri LuxR protein. J Bacteriol. 1992 Jun;174(12):4064–4069. doi: 10.1128/jb.174.12.4064-4069.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Choi S. H., Greenberg E. P. The C-terminal region of the Vibrio fischeri LuxR protein contains an inducer-independent lux gene activating domain. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11115–11119. doi: 10.1073/pnas.88.24.11115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Devine J. H., Shadel G. S., Baldwin T. O. Identification of the operator of the lux regulon from the Vibrio fischeri strain ATCC7744. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5688–5692. doi: 10.1073/pnas.86.15.5688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dunlap P. V., Greenberg E. P. Control of Vibrio fischeri luminescence gene expression in Escherichia coli by cyclic AMP and cyclic AMP receptor protein. J Bacteriol. 1985 Oct;164(1):45–50. doi: 10.1128/jb.164.1.45-50.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dunlap P. V., Greenberg E. P. Control of Vibrio fischeri lux gene transcription by a cyclic AMP receptor protein-luxR protein regulatory circuit. J Bacteriol. 1988 Sep;170(9):4040–4046. doi: 10.1128/jb.170.9.4040-4046.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Engebrecht J., Nealson K., Silverman M. Bacterial bioluminescence: isolation and genetic analysis of functions from Vibrio fischeri. Cell. 1983 Mar;32(3):773–781. doi: 10.1016/0092-8674(83)90063-6. [DOI] [PubMed] [Google Scholar]
  9. Engebrecht J., Silverman M. Identification of genes and gene products necessary for bacterial bioluminescence. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4154–4158. doi: 10.1073/pnas.81.13.4154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Engebrecht J., Silverman M. Nucleotide sequence of the regulatory locus controlling expression of bacterial genes for bioluminescence. Nucleic Acids Res. 1987 Dec 23;15(24):10455–10467. doi: 10.1093/nar/15.24.10455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fuqua C., Winans S. C., Greenberg E. P. Census and consensus in bacterial ecosystems: the LuxR-LuxI family of quorum-sensing transcriptional regulators. Annu Rev Microbiol. 1996;50:727–751. doi: 10.1146/annurev.micro.50.1.727. [DOI] [PubMed] [Google Scholar]
  12. Fuqua W. C., Winans S. C., Greenberg E. P. Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators. J Bacteriol. 1994 Jan;176(2):269–275. doi: 10.1128/jb.176.2.269-275.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gill S. C., von Hippel P. H. Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem. 1989 Nov 1;182(2):319–326. doi: 10.1016/0003-2697(89)90602-7. [DOI] [PubMed] [Google Scholar]
  14. Gray K. M., Passador L., Iglewski B. H., Greenberg E. P. Interchangeability and specificity of components from the quorum-sensing regulatory systems of Vibrio fischeri and Pseudomonas aeruginosa. J Bacteriol. 1994 May;176(10):3076–3080. doi: 10.1128/jb.176.10.3076-3080.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hanzelka B. L., Greenberg E. P. Evidence that the N-terminal region of the Vibrio fischeri LuxR protein constitutes an autoinducer-binding domain. J Bacteriol. 1995 Feb;177(3):815–817. doi: 10.1128/jb.177.3.815-817.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Henikoff S., Wallace J. C., Brown J. P. Finding protein similarities with nucleotide sequence databases. Methods Enzymol. 1990;183:111–132. doi: 10.1016/0076-6879(90)83009-x. [DOI] [PubMed] [Google Scholar]
  17. Hershberger C. D., Ye R. W., Parsek M. R., Xie Z. D., Chakrabarty A. M. The algT (algU) gene of Pseudomonas aeruginosa, a key regulator involved in alginate biosynthesis, encodes an alternative sigma factor (sigma E). Proc Natl Acad Sci U S A. 1995 Aug 15;92(17):7941–7945. doi: 10.1073/pnas.92.17.7941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ross W., Gosink K. K., Salomon J., Igarashi K., Zou C., Ishihama A., Severinov K., Gourse R. L. A third recognition element in bacterial promoters: DNA binding by the alpha subunit of RNA polymerase. Science. 1993 Nov 26;262(5138):1407–1413. doi: 10.1126/science.8248780. [DOI] [PubMed] [Google Scholar]
  19. Salmond G. P., Bycroft B. W., Stewart G. S., Williams P. The bacterial 'enigma': cracking the code of cell-cell communication. Mol Microbiol. 1995 May;16(4):615–624. doi: 10.1111/j.1365-2958.1995.tb02424.x. [DOI] [PubMed] [Google Scholar]
  20. Schaefer A. L., Val D. L., Hanzelka B. L., Cronan J. E., Jr, Greenberg E. P. Generation of cell-to-cell signals in quorum sensing: acyl homoserine lactone synthase activity of a purified Vibrio fischeri LuxI protein. Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9505–9509. doi: 10.1073/pnas.93.18.9505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Shadel G. S., Baldwin T. O. The Vibrio fischeri LuxR protein is capable of bidirectional stimulation of transcription and both positive and negative regulation of the luxR gene. J Bacteriol. 1991 Jan;173(2):568–574. doi: 10.1128/jb.173.2.568-574.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Shadel G. S., Devine J. H., Baldwin T. O. Control of the lux regulon of Vibrio fischeri. J Biolumin Chemilumin. 1990 Apr-Jun;5(2):99–106. doi: 10.1002/bio.1170050205. [DOI] [PubMed] [Google Scholar]
  23. Shadel G. S., Young R., Baldwin T. O. Use of regulated cell lysis in a lethal genetic selection in Escherichia coli: identification of the autoinducer-binding region of the LuxR protein from Vibrio fischeri ATCC 7744. J Bacteriol. 1990 Jul;172(7):3980–3987. doi: 10.1128/jb.172.7.3980-3987.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sitnikov D. M., Schineller J. B., Baldwin T. O. Transcriptional regulation of bioluminesence genes from Vibrio fischeri. Mol Microbiol. 1995 Sep;17(5):801–812. doi: 10.1111/j.1365-2958.1995.mmi_17050801.x. [DOI] [PubMed] [Google Scholar]
  25. Slock J., VanRiet D., Kolibachuk D., Greenberg E. P. Critical regions of the Vibrio fischeri luxR protein defined by mutational analysis. J Bacteriol. 1990 Jul;172(7):3974–3979. doi: 10.1128/jb.172.7.3974-3979.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Swartzman A., Kapoor S., Graham A. F., Meighen E. A. A new Vibrio fischeri lux gene precedes a bidirectional termination site for the lux operon. J Bacteriol. 1990 Dec;172(12):6797–6802. doi: 10.1128/jb.172.12.6797-6802.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Swift S., Throup J. P., Williams P., Salmond G. P., Stewart G. S. Quorum sensing: a population-density component in the determination of bacterial phenotype. Trends Biochem Sci. 1996 Jun;21(6):214–219. [PubMed] [Google Scholar]
  28. Ulitzur S., Kuhn J. The transcription of bacterial luminescence is regulated by sigma 32. J Biolumin Chemilumin. 1988 Apr-Jun;2(2):81–93. doi: 10.1002/bio.1170020205. [DOI] [PubMed] [Google Scholar]
  29. Ulitzur S. The regulatory control of the bacterial luminescence system--a new view. J Biolumin Chemilumin. 1989 Jul;4(1):317–325. doi: 10.1002/bio.1170040144. [DOI] [PubMed] [Google Scholar]
  30. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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