1887

Abstract

and are capable of forming mixed biofilms in the lungs of cystic fibrosis patients. Both bacteria employ quorum-sensing systems, which rely on -acylhomoserine lactone (AHL) signal molecules, to co-ordinate expression of virulence factors with the formation of biofilms. As both bacteria utilize the same class of signal molecules the authors investigated whether communication between the species occurs. To address this issue, novel Gfp-based biosensors for non-destructive, detection of AHLs were constructed and characterized. These sensors were used to visualize AHL-mediated communication in mixed biofilms, which were cultivated either in artificial flow chambers or in alginate beads in mouse lung tissue. In both model systems was capable of perceiving the AHL signals produced by , while the latter strain did not respond to the molecules produced by . Measurements of extracellular proteolytic activities of defined quorum-sensing mutants grown in media complemented with AHL extracts prepared from culture supernatants of various wild-type and mutant strains supported the view of unidirectional signalling between the two strains.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-147-12-3249
2001-12-01
2019-11-14
Loading full text...

Full text loading...

/deliver/fulltext/micro/147/12/1473249a.html?itemId=/content/journal/micro/10.1099/00221287-147-12-3249&mimeType=html&fmt=ahah

References

  1. Andersen, J. B., Sternberg, C., Poulsen, L. K., Bjorn, S. P., Givskov, M. & Molin, S. ( 1998; ). New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria. Appl Environ Microbiol 64, 2240-2246.
    [Google Scholar]
  2. Andersen, J. B., Heydorn, A., Hentzer, M., Eberl, L., Geisenberger, O., Christensen, B. B., Molin, S. & Givskov, M. ( 2001; ). gfp based N-acyl-homoserine-lactone monitors for detection of bacterial communication. Appl Environ Microbiol 67, 575-585.[CrossRef]
    [Google Scholar]
  3. Ayora, S. & Götz, F. ( 1994; ). Genetic and biochemical properties of an extracellular neutral metalloprotease from Staphylococcus hyicus subsp. hyicus. Mol Gen Genet 242, 421-430.
    [Google Scholar]
  4. Bertani, G. ( 1951; ). Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol 62, 293-300.
    [Google Scholar]
  5. Christensen, B. B., Sternberg, C., Andersen, J. B., Palmer, R. J.Jr, Nielsen, A. T., Givskov, M. & Molin, S. ( 1999; ). Molecular tools for study of biofilm physiology. Methods Enzymol 310, 20-42.
    [Google Scholar]
  6. Clark, D. J. & Maaløe, O. ( 1967; ). DNA replication and the division cycle in Escherichia coli. J Mol Biol 23, 99-112.[CrossRef]
    [Google Scholar]
  7. Davies, G. D., Parsek, M. R., Pearson, J. P., Iglewski, B. H., Costeron, J. W. & Greenberg, E. P. ( 1998; ). The involvement of cell-to-cell signals in the development of a bacterial biofilm. Science 280, 295-298.[CrossRef]
    [Google Scholar]
  8. Eberl, L. ( 1999; ). N-Acyl homoserinelactone-mediated gene regulation in gram-negative bacteria. Syst Appl Microbiol 22, 493-506.[CrossRef]
    [Google Scholar]
  9. Eberl, L., Molin, S. & Givskov, M. ( 1999; ). Surface motility of Serratia liquefaciens MG1. J Bacteriol 181, 1703-1712.
    [Google Scholar]
  10. Evans, K., Passador, L., Srikumar, R., Tsang, E., Nezezon, J. & Poole, K. ( 1998; ). Influence of the MexAB-OprM multidrug efflux system on quorum sensing in Pseudomonas aeruginosa. J Bacteriol 180, 5443-5447.
    [Google Scholar]
  11. Fuqua, C., Winans, S. C. & Greenberg, E. P. ( 1996; ). Census and consensus in bacterial ecosystems: the LuxR-LuxI family of quorum-sensing transcriptional regulators. Annu Rev Microbiol 50, 727-751.[CrossRef]
    [Google Scholar]
  12. Geisenberger, O., Givskov, M., Riedel, K., Høiby, N., Tümmler, B. & Eberl, L. ( 2000; ). Production of N-acyl-l-homoserine lactones by P. aeruginosa isolates from chronic lung infections associated with cystic fibrosis. FEMS Microbiol Lett 184, 273-278.
    [Google Scholar]
  13. Gotschlich, A., Huber, B., Geisenberger, O. & 11 other authors ( 2001; ). Synthesis of multiple N-acyl-homoserine lactones is wide-spread among members of the Burkholderia cepacia complex. Syst Appl Microbiol 24, 1–14.[CrossRef]
    [Google Scholar]
  14. Govan, J. R. W. & Deretic, V. ( 1996; ). Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia. Microbiol Rev 60, 539-574.
    [Google Scholar]
  15. Govan, J. R. W., Hughes, J. E. & Vandamme, P. ( 1996; ). Burkholderia cepacia: medical, taxonomic and ecological issues. J Med Microbiol 45, 395-407.[CrossRef]
    [Google Scholar]
  16. Hardman, A. M., Stewart, G. S. & Williams, P. ( 1998; ). Quorum sensing and the cell-cell communication dependent regulation of gene expression in pathogenic and non-pathogenic bacteria. Antonie Leeuwenhoek 74, 199-210.[CrossRef]
    [Google Scholar]
  17. Hentzer, M., Riedel, K., Rasmussen, T. B. & 9 other authors (2002). Inhibition of quorum sensing in Pseudomonas aeruginosa biofilm bacteria by a halogenated furanone compound. Microbiology 148 (in press).
  18. Herrero, M., de Lorenzo, V. & 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]
  19. Holloway, B. W. ( 1955; ). Genetic recombination in Pseudomonas aeruginosa. J Gen Microbiol 13, 572-581.[CrossRef]
    [Google Scholar]
  20. Huber, B., Riedel, K., Hentzer, M., Heydorn, A., Gotschlich, A., Givskov, M., Molin, S. & Eberl, L. ( 2001; ). The cep quorum-sensing system of Burkholderia cepacia H111 controls biofilm formation and swarming motility. Microbiology 147, 2517-2528.
    [Google Scholar]
  21. Isles, A., Maclusky, I., Corey, M., Gold, R., Prober, C., Fleming, P. & Levison, H. ( 1984; ). Pseudomonas cepacia infection in cystic fibrosis: an emerging problem. J Pediatr 104, 206-210.[CrossRef]
    [Google Scholar]
  22. Johansen, H. K., Espersen, F., Pedersen, S. S., Hougen, H. P., Rygaard, J. & Høiby, N. ( 1993; ). Chronic Pseudomonas aeruginosa lung infection in normal and athymic rats. APMIS 101, 207-225.[CrossRef]
    [Google Scholar]
  23. Kessler, B., de Lorenzo, V. & Timmis, K. N. ( 1992; ). A general system to integrate lacZ fusions into the chromosomes of gram-negative eubacteria: regulation of the Pm promoter of the TOL plasmid studied with all controlling elements in monocopy. Mol Gen Genet 233, 293-301.[CrossRef]
    [Google Scholar]
  24. de Kievit, T. R. & Iglewski, B. H. ( 2000; ). Bacterial quorum sensing in pathogenic relationships. Infect Immun 68, 4839-4849.[CrossRef]
    [Google Scholar]
  25. Kovach, M. E., Elzer, P. H., Hill, D. S., Robertson, G. T., Farris, M. A., Roop, R. M.II & Peterson, K. M. ( 1995; ). Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene 166, 175-176.[CrossRef]
    [Google Scholar]
  26. Lewenza, S., Conway, B., Greenberg, E. P. & Sokol, P. A. ( 1999; ). Quorum sensing in Burkholderia cepacia: identification of the LuxRI homologs CepIR. J Bacteriol 181, 748-756.
    [Google Scholar]
  27. McClean, K. H., Winson, M. K., Fish, L. & 9 other authors ( 1997; ). Quorum sensing and Chromobacterium violaceum: exploitation of violacein production and inhibition for the detection of N-acylhomoserine lactones. Microbiology 143, 3703–3711.[CrossRef]
    [Google Scholar]
  28. McKenney, D., Brown, K. E. & Allison, D. G. ( 1995; ). Influence of Pseudomonas aeruginosa exoproducts on virulence factor production in Burkholderia cepacia: evidence of interspecies communication. J Bacteriol 177, 6989-6992.
    [Google Scholar]
  29. Møller, S., Sternberg, C., Andersen, J. B., Christensen, B. B., Ramos, J. L., Givskov, M. & Molin, S. ( 1998; ). In situ gene expression in mixed-culture biofilms: evidence of metabolic interactions between community members. Appl Environ Microbiol 64, 721-732.
    [Google Scholar]
  30. Moser, C., Johansen, H. K., Song, Z. J., Hougen, H. P., Rygaard, J. & Høiby, N. ( 1997; ). Chronic Pseudomonas aeruginosa lung infection is more severe in Th2 responding BALB/c mice compared to Th1 responding C3H/HeN mice. APMIS 105, 838-842.[CrossRef]
    [Google Scholar]
  31. Parsek, M. R. & Greenberg, E. P. ( 1999; ). Quorum sensing signals in development of Pseudomonas aeruginosa biofilms. Methods Enzymol 310, 43-55.
    [Google Scholar]
  32. Passador, L., Cook, J. M., Gambello, M. J., Rust, L. & Iglewski, B. H. ( 1993; ). Expression of Pseudomonas aeruginosa virulence genes requires cell-to-cell communication. Science 260, 1127-1130.[CrossRef]
    [Google Scholar]
  33. Pearson, J. P., Gray, K. M., Passador, L., Tucker, K. D., Eberhard, A., Iglewski, B. H. & Greenberg, E. P. ( 1994; ). Structure of the autoinducer required for expression of Pseudomonas aeruginosa virulence genes. Proc Natl Acad Sci USA 91, 197-201.[CrossRef]
    [Google Scholar]
  34. Pearson, J. P., Van Delden, C. & Iglewski, B. H. ( 1999; ). Active efflux and diffusion are involved in transport of Pseudomonas aeruginosa cell-to-cell signals. J Bacteriol 181, 1203-1210.
    [Google Scholar]
  35. Pesci, E. C., Pearson, J. P., Seed, P. C. & Iglewski, B. H. ( 1997; ). Regulation of las and rhl quorum sensing in Pseudomonas aeruginosa. J Bacteriol 179, 3127-3132.
    [Google Scholar]
  36. Römling, U., Fiedler, B., Bosshammer, J., Grothues, D., Greipel, J., von der Hardt, H. & Tümmler, B. ( 1994; ). Epidemiology of chronic Pseudomonas aeruginosa infections in cystc fibrosis. J Infect Dis 170, 1617-1621.
    [Google Scholar]
  37. Rumbaugh, K. P., Griswold, J. A., Iglewski, B. H. & Hamood, A. N. ( 1999; ). Contribution of quorum sensing to the virulence of Pseudomonas aeruginosa in burn wound infections. Infect Immun 67, 5854-5862.
    [Google Scholar]
  38. Saiman, L., Cacalano, G. & Prince, A. ( 1990; ). Pseudomonas cepacia adherence to respiratory epithelial cells is enhanced by Pseudomonas aeruginosa. Infect Immun 58, 2578-2584.
    [Google Scholar]
  39. Salmond, G. P., Bycroft, B. W., Stewart, G. S. & Williams, P. ( 1995; ). The bacterial ‘enigma’: cracking the code of cell-cell communication. Mol Microbiol 16, 615-624.[CrossRef]
    [Google Scholar]
  40. Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  41. Schierholz, J. M., Beuth, J., König, D., Nürnberger, A. & Pulverer, G. ( 1999; ). Antimicrobial substances and effects on sessile bacteria. Zentbl Bakteriol 289, 165-177.[CrossRef]
    [Google Scholar]
  42. Singh, P. K., Schaefer, A. L., Parsek, M. R., Moninger, T. O., Welsh, M. J. & Greenberg, E. P. ( 2000; ). Quorum-sensing signals indicate that cystic fibrosis lungs are infected with bacterial biofilms. Nature 407, 762-764.[CrossRef]
    [Google Scholar]
  43. Swift, S., Throup, J. P., Williams, P., Salmond, G. P. & Stewart, G. S. ( 1994; ). Quorum sensing: a population-density component in the determination of bacterial phenotype. Trends Biochem Sci 21, 214-219.
    [Google Scholar]
  44. Tan, M. W., Mahajan-Miklos, S. & Ausubel, F. M. ( 1999; ). Killing of Caenorhabditis elegans by Pseudomonas aeruginosa used to model mammalian bacterial pathogenesis. Proc Natl Acad Sci USA 96, 715-70.[CrossRef]
    [Google Scholar]
  45. Tang, H. B., DiMango, E., Bryan, R., Gambello, M., Iglewski, B. H., Goldberg, J. B. & Prince, A. ( 1996; ). Contribution of specific Pseudomonas aeruginosa virulence factors to pathogenesis of pneumonia in a neonatal mouse model of infection. Infect Immun 64, 37-43.
    [Google Scholar]
  46. Tümmler, B. & Kiewitz, C. ( 1999; ). Cystic fibrosis: an inherited susceptibility to bacterial respiratory infections. Mol Med Today 5, 351-358.[CrossRef]
    [Google Scholar]
  47. Van Delden, C. & Iglewski, B. ( 1998; ). Cell-to-cell signaling and Pseudomonas aeruginosa infections. Emerg Infect Dis 4, 551-560.[CrossRef]
    [Google Scholar]
  48. Williams, P., Camara, M., Hardman, A. & 7 other authors ( 2000; ). Quorum sensing and the population-dependent control of virulence. Philos Trans R Soc Lond B Biol Sci 355, 667–680.[CrossRef]
    [Google Scholar]
  49. Winson, M. K., Camara, M., Lafiti, A. & 10 other authors ( 1995; ). Multiple N-acyl-l-homoserine lactone signal molecules regulate production of virulence determinants and secondary metabolites in Pseudomonas aeruginosa. Proc Natl Acad Sci USA 92, 9427–9431.[CrossRef]
    [Google Scholar]
  50. Winson, M. K., Swift, S., Fish, L., Throup, J. P., Jorgensen, F., Chhabra, S. R., Bycroft, B. W., Williams, P. & Stewart, G. S. A. B. ( 1998; ). Construction and analysis of luxCDABE-based plasmid sensors for investigating N-acyl-homoserine lactone-mediated quorum sensing. FEMS Microbiol Lett 163, 185-192.[CrossRef]
    [Google Scholar]
  51. Wu, H., Song, Z., Hentzer, M. & 8 other authors ( 2000; ). Detection of N-acylhomoserine lactones in lung tissues of mice infected with Pseudomonas aeruginosa. Microbiology 146, 2481–2493.
    [Google Scholar]
  52. Xu, K. D., McFeter, G. A. & Stewart, P. S. ( 2000; ). Biofilm resistance to antimicrobial agents. Microbiology 146, 547-549.
    [Google Scholar]
  53. Zhu, J., Beaber, J. W., Moré, M. I., Fuqua, C., Eberhard, A. & Winans, S. C. ( 1998; ). Analogs of the autoinducer 3-oxooctanoyl-homoserine lactone strongly inhibit activity of the TraR protein of Agrobacterium tumefaciens. J Bacteriol 180, 5398-5405.
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-147-12-3249
Loading
/content/journal/micro/10.1099/00221287-147-12-3249
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