1887

Abstract

A quorum-sensing locus, /, which is involved in the regulation of root colonization and plant disease-suppressive ability, was previously identified in 2P24. In this study, we performed random mutagenesis using mini-Tn in order to screen the upstream transcriptional regulators of , a biosynthase gene responsible for the synthesis of -acylhomoserine lactone signal molecules. Two mutants, PM400 and PM410, with elevated gene promoter activity, were identified from ∼10 000 insertion clones. The amino acid sequences of the interrupted genes in these two mutants were highly similar to PhoQ, a sensor protein of the two-component regulatory system PhoP/PhoQ, which responds to environmental Mg starvation and regulates virulence in and antimicrobial peptide resistance in . The promoter activity of was also induced under low-Mg conditions in the 2P24 strain of . Deletion mutagenesis and complementation experiments demonstrated that the transcription of was negatively regulated by the sensor PhoQ but positively regulated by the response regulator PhoP. Genetic evidence also indicated that transcription of the outer-membrane protein gene was induced by Mg starvation through regulation of the wild-type PhoP/PhoQ system. Additionally, PhoQ was involved in biofilm formation by 2P24 under low-Mg conditions through a PhoP-independent pathway.

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2009-01-01
2019-09-24
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References

  1. Bertani, I. & Venturi, V. ( 2004; ). Regulation of the N-acyl homoserine lactone-dependent quorum-sensing system in rhizosphere Pseudomonas putida WCS358 and cross-talk with the stationary-phase RpoS sigma factor and the global regulator GacA. Appl Environ Microbiol 70, 5493–5502.[CrossRef]
    [Google Scholar]
  2. Brinkman, F. S., Macfarlane, E. L., Warrener, P. & Hancock, R. E. ( 2001; ). Evolutionary relationships among virulence-associated histidine kinases. Infect Immun 69, 5207–5211.[CrossRef]
    [Google Scholar]
  3. Castelli, M. E., Ve'scovi, E. G. & Soncini, F. C. ( 2000; ). The phosphatase activity is the target for Mg2+ regulation of the sensor protein PhoQ in Salmonella. J Biol Chem 275, 22948–22954.[CrossRef]
    [Google Scholar]
  4. Cha, C., Gao, P., Chen, Y. C., Shaw, P. D. & Farrand, S. K. ( 1998; ). Production of acyl-homoserine lactone quorum-sensing signals by Gram-negative plant-associated bacteria. Mol Plant Microbe Interact 11, 1119–1129.[CrossRef]
    [Google Scholar]
  5. Chilton, M. D., Currier, T. C., Farrand, S. K., Bendich, A. J., Gordon, M. P. & Nester, E. W. ( 1974; ). Agrobacterium tumefaciens DNA and PS8 bacteriophage DNA not detected in crown gall tumors. Proc Natl Acad Sci U S A 71, 3672–3676.[CrossRef]
    [Google Scholar]
  6. de Kievit, T. R. & Iglewski, B. H. ( 2000; ). Bacterial quorum sensing in pathogenic relationships. Infect Immun 68, 4839–4849.[CrossRef]
    [Google Scholar]
  7. Eguchi, Y., Itou, J., Yamane, M., Demizu, R., Yamato, F., Okada, A., Mori, H., Kato, A. & Utsumi, R. ( 2007; ). B1500, a small membrane protein, connects the two-component systems EvgS/EvgA and PhoQ/PhoP in Escherichia coli. Proc Natl Acad Sci U S A 104, 18712–18717.[CrossRef]
    [Google Scholar]
  8. Ernst, R. K., Yi, E. C., Guo, L., Lim, K. B., Burns, J. L., Hackett, M. & Miller, S. I. ( 1999; ). Specific lipopolysaccharide found in cystic fibrosis airway Pseudomonas aeruginosa. Science 286, 1561–1565.[CrossRef]
    [Google Scholar]
  9. González, J. E. & Keshavan, N. D. ( 2006; ). Messing with bacterial quorum sensing. Microbiol Mol Biol Rev 70, 859–875.[CrossRef]
    [Google Scholar]
  10. Groisman, E. A. ( 2001; ). The pleiotropic two-component regulatory system PhoP-PhoQ. J Bacteriol 183, 1835–1842.[CrossRef]
    [Google Scholar]
  11. Herrero, M., Lorenzo, V. D. & Timmis, K. N. ( 1990; ). Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in gram-negative bacteria. J Bacteriol 172, 6557–6567.
    [Google Scholar]
  12. Jensen, V., Löns, D., Zaoui, C., Bredenbruch, F., Meissner, A., Dieterich, G., Münch, R. & Häussler, S. ( 2006; ). RhlR expression in Pseudomonas aeruginosa is modulated by the Pseudomonas quinolone signal via PhoB-dependent and -independent pathways. J Bacteriol 188, 8601–8606.[CrossRef]
    [Google Scholar]
  13. Juhas, M., Wiehlmann, L., Huber, B., Jordan, D., Lauber, J., Salunkhe, P., Limpert, A. S., von Goõta, F., Steinmetz, I. & other authors ( 2004; ). Global regulation of quorum sensing and virulence by VqsR in Pseudomonas aeruginosa. Microbiology 150, 831–841.[CrossRef]
    [Google Scholar]
  14. Kasahara, M., Nakata, A. & Shinagawa, H. ( 1992; ). Molecular analysis of the Escherichia coli PhoP-PhoQ operon. J Bacteriol 174, 492–498.
    [Google Scholar]
  15. Kato, A., Tanabe, H. & Utsumi, R. ( 1999; ). Molecular characterization of the PhoP-PhoQ two-component system in Escherichia coli K-12: identification of extracellular Mg2+-responsive promoters. J Bacteriol 181, 5516–5520.
    [Google Scholar]
  16. Keen, N. T., Tamaki, S., Kobayashi, D. & Trollinger, D. ( 1988; ). Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria. Gene 70, 191–197.[CrossRef]
    [Google Scholar]
  17. Keller, L. & Surette, M. G. ( 2006; ). Communication in bacteria: an ecological and evolutionary perspective. Nat Rev Microbiol 4, 249–258.[CrossRef]
    [Google Scholar]
  18. Kier, L. D., Weppelman, R. M. & Ames, B. N. ( 1979; ). Regulation of nonspecific acid phosphatase in Salmonella: phoN and phoP genes. J Bacteriol 138, 155–161.
    [Google Scholar]
  19. Macfarlane, E. L., Kwasnicka, A., Ochs, M. M. & Hancock, R. E. ( 1999; ). PhoP-PhoQ homologues in Pseudomonas aeruginosa regulate expression of the outer-membrane protein OprH and polymyxin B resistance. Mol Microbiol 34, 305–316.[CrossRef]
    [Google Scholar]
  20. Macfarlane, E. L. A., Kwasnicka, A. & Hancock, R. E. W. ( 2000; ). Role of Pseudomonas aeruginosa in resistance to antimicrobial peptides and aminoglycosides. Microbiology 146, 2543–2554.
    [Google Scholar]
  21. Matsubara, M., Kitaota, S., Takeda, S. & Mizuno, T. ( 2000; ). Tuning of the porin expression under anaerobic growth conditions by His-to-Asp cross-phosphorelay through both the EnvZ-osmosensor and ArcB-anaerosensor in Escherichia coli. Genes Cells 5, 555–569.[CrossRef]
    [Google Scholar]
  22. McPhee, J. B., Bains, M., Winsor, G., Lewenza, S., Kwasnicka, A., Brazas, M. D., Brinkman, F. S. & Hancock, R. E. ( 2006; ). Contribution of the PhoP-PhoQ and PmrA-PmrB two-component regulatory systems to Mg2+-induced gene regulation in Pseudomonas aeruginosa. J Bacteriol 188, 3995–4006.[CrossRef]
    [Google Scholar]
  23. Miller, J. H. ( 1972; ). Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  24. Miller, M. B. & Bassler, B. L. ( 2001; ). Quorum sensing in bacteria. Annu Rev Microbiol 55, 165–199.[CrossRef]
    [Google Scholar]
  25. Miller, S. I., Kukral, A. M. & Mekalanos, J. J. ( 1989; ). A two-component regulatory system (phoP phoQ) controls Salmonella typhimurium virulence. Proc Natl Acad Sci U S A 86, 5054–5058.[CrossRef]
    [Google Scholar]
  26. Minagawa, S., Ogasawara, H., Kato, A., Yamamoto, K., Eguchi, Y., Oshima, T., Mori, H., Ishihama, A. & Utsumi, R. ( 2003; ). Identification and molecular characterization of the Mg2+ stimulon of Escherichia coli. J Bacteriol 185, 3696–3702.[CrossRef]
    [Google Scholar]
  27. Pessi, G., Williams, F., Hindle, Z., Heurlier, K., Holden, M. T., Camara, M., Haas, D. & Williams, P. ( 2001; ). The global posttranscriptional regulator RsmA modulates production of virulence determinants and N-acylhomoserine lactones in Pseudomonas aeruginosa. J Bacteriol 183, 6676–6683.[CrossRef]
    [Google Scholar]
  28. Prouty, A. M. & Gunn, J. S. ( 2003; ). Comparative analysis of Salmonella enterica serovar Typhimurium biofilm formation on gallstones and on glass. Infect Immun 71, 7154–7158.[CrossRef]
    [Google Scholar]
  29. Rampioni, G., Polticelli, F., Bertani, I., Righetti, K., Venturi, V., Zennaro, E. & Leoni, L. ( 2007; ). The Pseudomonas quorum-sensing regulator RsaL belongs to the tetrahelical superclass of H-T-H proteins. J Bacteriol 189, 1922–1930.[CrossRef]
    [Google Scholar]
  30. Ramsey, M. M. & Whiteley, M. ( 2004; ). Pseudomonas aeruginosa attachment and biofilm development in dynamic environments. Mol Microbiol 53, 1075–1087.[CrossRef]
    [Google Scholar]
  31. Reimmann, C., Beyeler, M., Latifi, A., Winteler, H., Foglino, M., Lazdunski, A. & Haas, D. ( 1997; ). The global activator GacA of Pseudomonas aeruginosa PAO1 positively controls the production of the autoinducer N-butyryl-homoserine lactone and the formation of the virulence factors pyocyanin, cyanide, and lipase. Mol Microbiol 24, 309–319.[CrossRef]
    [Google Scholar]
  32. Sakuragi, Y. & Kolter, R. ( 2007; ). Quorum-sensing regulation of the biofilm matrix genes (pel) of Pseudomonas aeruginosa. J Bacteriol 189, 5383–5386.[CrossRef]
    [Google Scholar]
  33. Sambrook, J., Fritsch, E. F. & Maniatis, T. ( 1989; ). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  34. Soncini, F. C., Véscovi, E. G., Solomon, F. & Groisman, E. A. ( 1996; ). Molecular basis of the magnesium deprivation response in Salmonella typhimurium: identification of PhoP-regulated genes. J Bacteriol 178, 5092–5099.
    [Google Scholar]
  35. Van den Eede, G., Deblaere, R., Goethals, K., Van Montagu, M. & Holsters, M. ( 1992; ). Broad host range and promoter selection vectors for bacteria that interact with plants. Mol Plant Microbe Interact 5, 228–234.[CrossRef]
    [Google Scholar]
  36. Venturi, V. ( 2006; ). Regulation of quorum sensing in Pseudomonas. FEMS Microbiol Rev 30, 274–291.[CrossRef]
    [Google Scholar]
  37. Véscovi, E. G., Soncini, F. C. & Groisman, E. G. ( 1996; ). Mg2+ as an extracellular signal: environmental regulation of Salmonella virulence. Cell 84, 165–174.[CrossRef]
    [Google Scholar]
  38. Waters, C. M., Lu, W., Rabinowitz, J. D. & Bassler, B. L. ( 2008; ). Quorum sensing controls biofilm formation in Vibrio cholerae through modulation of cyclic di-GMP levels and repression of vpsT. J Bacteriol 190, 2527–2536.[CrossRef]
    [Google Scholar]
  39. Wei, H. L. & Zhang, L. Q. ( 2005; ). Cloning and functional characterization of the gacS gene of the biocontrol strain Pseuodomonas fluorescens 2P24. Wei Sheng Wu Xue Bao 45, 368–372.
    [Google Scholar]
  40. Wei, H. L. & Zhang, L. Q. ( 2006; ). Quorum-sensing system influences root colonization and biological control ability in Pseudomonas fluorescens 2P24. Antonie Van Leeuwenhoek 89, 267–280.[CrossRef]
    [Google Scholar]
  41. Yan, Q., Wu, X. G., Wei, H. L., Wang, H. M. & Zhang, L. Q. ( 2008; ). Differential control of the PcoI-PcoR quorum-sensing system in Pseudomonas fluorescens 2P24 by sigma factor RpoS and the GacS/GacA two-component regulatory system. Microbiol Res
    [Google Scholar]
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Detailed protocol for construction of and in-frame deletion mutants in strain 2P24 [ PDF] (52 kb) Primers used in this study [ PDF] (14 kb) Mg concentration regulated expression in strain 2P24 [ PDF] (51 kb) Alignment of the PhoP and the PhoQ proteins [ PDF] (141 kb) Mg regulates expression in 2P24 [ PDF] (20 kb) Influence of PhoP/PhoQ on growth properties in LB medium [ PDF] (22 kb)

PDF

Detailed protocol for construction of and in-frame deletion mutants in strain 2P24 [ PDF] (52 kb) Primers used in this study [ PDF] (14 kb) Mg concentration regulated expression in strain 2P24 [ PDF] (51 kb) Alignment of the PhoP and the PhoQ proteins [ PDF] (141 kb) Mg regulates expression in 2P24 [ PDF] (20 kb) Influence of PhoP/PhoQ on growth properties in LB medium [ PDF] (22 kb)

PDF

Detailed protocol for construction of and in-frame deletion mutants in strain 2P24 [ PDF] (52 kb) Primers used in this study [ PDF] (14 kb) Mg concentration regulated expression in strain 2P24 [ PDF] (51 kb) Alignment of the PhoP and the PhoQ proteins [ PDF] (141 kb) Mg regulates expression in 2P24 [ PDF] (20 kb) Influence of PhoP/PhoQ on growth properties in LB medium [ PDF] (22 kb)

PDF

Detailed protocol for construction of and in-frame deletion mutants in strain 2P24 [ PDF] (52 kb) Primers used in this study [ PDF] (14 kb) Mg concentration regulated expression in strain 2P24 [ PDF] (51 kb) Alignment of the PhoP and the PhoQ proteins [ PDF] (141 kb) Mg regulates expression in 2P24 [ PDF] (20 kb) Influence of PhoP/PhoQ on growth properties in LB medium [ PDF] (22 kb)

PDF

Detailed protocol for construction of and in-frame deletion mutants in strain 2P24 [ PDF] (52 kb) Primers used in this study [ PDF] (14 kb) Mg concentration regulated expression in strain 2P24 [ PDF] (51 kb) Alignment of the PhoP and the PhoQ proteins [ PDF] (141 kb) Mg regulates expression in 2P24 [ PDF] (20 kb) Influence of PhoP/PhoQ on growth properties in LB medium [ PDF] (22 kb)

PDF

Detailed protocol for construction of and in-frame deletion mutants in strain 2P24 [ PDF] (52 kb) Primers used in this study [ PDF] (14 kb) Mg concentration regulated expression in strain 2P24 [ PDF] (51 kb) Alignment of the PhoP and the PhoQ proteins [ PDF] (141 kb) Mg regulates expression in 2P24 [ PDF] (20 kb) Influence of PhoP/PhoQ on growth properties in LB medium [ PDF] (22 kb)

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