1887

Abstract

Extracellular DNA is one of the major matrix components in biofilms. It functions as an intercellular connector and plays a role in stabilization of the biofilms. Evidence that DNA release in PAO1 biofilms is controlled by the - and quorum-sensing systems has been previously presented. This paper provides evidence that DNA release in PAO1 biofilms is also under iron regulation. Experiments involving cultivation of in microtitre trays suggested that expression, DNA release and biofilm formation were favoured in media with low iron concentrations (5 μM FeCl), and decreased with increasing iron concentrations. Experiments involving cultivation of in a flow-chamber system suggested that a high level of iron (100 μM FeCl) in the medium suppressed DNA release, structural biofilm development, and the development of subpopulations with increased tolerance toward antimicrobial compounds. Experiments with strains harbouring fluorescent reporters suggested that expression of the operon was induced in particular subpopulations of the biofilm cells under low-iron conditions (1 μM FeCl), but repressed in the biofilm cells under high-iron conditions (100 μM FeCl).

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2007-05-01
2020-08-06
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References

  1. Allesen-Holm M., Barken K. B., Yang L., Klausen M., Webb J. S., Kjelleberg S., Molin S., Givskov M., Tolker-Nielsen T.. 2006; A characterization of DNA release in Pseudomonas aeruginosa cultures and biofilms. Mol Microbiol59:1114–1128[CrossRef]
    [Google Scholar]
  2. 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 Microbiol64:2240–2246
    [Google Scholar]
  3. 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 sensor systems for detection of bacterial communication. Appl Environ Microbiol67:575–585[CrossRef]
    [Google Scholar]
  4. Baltimore R. S., Christie C. D., Smith G. J.. 1989; Immunohistopathologic localization of Pseudomonas aeruginosa in lungs from patients with cystic fibrosis. Implications for the pathogenesis of progressive lung deterioration. Am Rev Respir Dis140:1650–1661[CrossRef]
    [Google Scholar]
  5. Banin E., Vasil M. L., Greenberg E. P.. 2005; Iron and Pseudomonas aeruginosa biofilm formation. Proc Natl Acad Sci U S A102:11076–11081[CrossRef]
    [Google Scholar]
  6. Banin E., Brady K. M., Greenberg E. P.. 2006; Chelator-induced dispersal and killing of Pseudomonas aeruginosa cells in a biofilm. App Environ Microbial72:2064–2069[CrossRef]
    [Google Scholar]
  7. Bertani G.. 1951; Studies on lysogenesis. I. The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol62:293–300
    [Google Scholar]
  8. Bjarnsholt T., Jensen P. O., Burmolle M., Hentzer M., Haagensen J. A., Hougen H. P., Calum H., Madsen K. G., Moser C.. other authors 2005; Pseudomonas aeruginosa tolerance to tobramycin, hydrogen peroxide and polymorphonuclear leukocytes is quorum-sensing dependent. Microbiology151:373–383[CrossRef]
    [Google Scholar]
  9. Bjorn M. J., Sokol P. A., Iglewski B. H.. 1979; Influence of iron on yields of extracellular products in Pseudomonas aeruginosa cultures. J Bacteriol138:193–200
    [Google Scholar]
  10. Bollinger N., Hassett D. J., Iglewski B. H., Costerton J. W., McDermott T. R.. 2001; Gene expression in Pseudomonas aeruginosa : evidence of iron override effects on quorum sensing and biofilm-specific gene regulation. J Bacteriol183:1990–1996[CrossRef]
    [Google Scholar]
  11. Brint J. M., Ohman D. E.. 1995; Synthesis of multiple exoproducts in Pseudomonas aeruginosa is under the control of RhlR-RhlI, another set of regulators in strain PAO1 with homology to the autoinducer-responsive LuxR-LuxI family. J Bacteriol177:7155–7163
    [Google Scholar]
  12. Cao H., Krishnan G., Goumnerov B., Tsongalis J., Tompkins R., Rahme L. G.. 2001; A quorum sensing-associated virulence gene of Pseudomonas aeruginosa encodes a LysR-like transcription regulator with a unique self-regulatory mechanism. Proc Natl Acad Sci U S A98:14613–14618[CrossRef]
    [Google Scholar]
  13. Clark D. J., Maaløe O.. 1967; DNA replication and the division cycle in Escherichia coli. J Mol Biol23:99–112[CrossRef]
    [Google Scholar]
  14. Cornelis P., Aendekerk S.. 2004; A new regulator linking quorum sensing and iron uptake in Pseudomonas aeruginosa. Microbiology150:752–756[CrossRef]
    [Google Scholar]
  15. Costerton J. W., Stewart P. S., Greenberg E. P.. 1999; Bacterial biofilms: a common cause of persistent infections. Science284:1318–1322[CrossRef]
    [Google Scholar]
  16. D'Argenio D. A., Calfee M. W., Rainey P. B., Pesci E. C.. 2002; Autolysis and autoaggregation in Pseudomonas aeruginosa colony morphology mutants. J Bacteriol184:6481–6489[CrossRef]
    [Google Scholar]
  17. Davey M. E., O'Toole G. A.. 2000; Microbial biofilms: from ecology to molecular genetics. Microbiol Mol Biol Rev64:847–867[CrossRef]
    [Google Scholar]
  18. Davies D. G., Parsek M. R., Pearson J. P., Iglewski B. H., Costerton J. W., Greenberg E. P.. 1998; The involvement of cell-to-cell signals in the development of a bacterial biofilm. Science280:295–298[CrossRef]
    [Google Scholar]
  19. Gallagher L. A., McKnight S. L., Kuznetsova M. S., Pesci E. C., Manoil C.. 2002; Functions required for extracellular quinolone signaling by Pseudomonas aeruginosa. J Bacteriol184:6472–6480[CrossRef]
    [Google Scholar]
  20. Haagensen J. A., Klausen M., Ernst R. K., Miller S. I., Folkesson A., Tolker-Nielsen T., Molin S.. 2007; Differentiation and distribution of colistin/SDS tolerant cells in Pseudomonas aeruginosa biofilms. J Bacteriol189:28–37[CrossRef]
    [Google Scholar]
  21. Haas B., Kraut J., Marks J., Zanker S. C., Castignetti D.. 1991; Siderophore presence in sputa of cystic fibrosis patients. Infect Immun59:3997–4000
    [Google Scholar]
  22. Hassett D. J., Charniga L., Bean K., Ohman D. E., Cohen M. S.. 1992; Response of Pseudomonas aeruginosa to pyocyanin: mechanisms of resistance, antioxidant defenses, and demonstration of a manganese-cofactored superoxide dismutase. Infect Immun60:328–336
    [Google Scholar]
  23. Hassett D. J., Howell M. L., Sokol P. A., Vasil M. L., Dean G. E.. 1997; Fumarase C activity is elevated in response to iron deprivation and in mucoid, alginate-producing Pseudomonas aeruginosa : cloning and characterization of fumC and purification of native fumC. J Bacteriol179:1442–1451
    [Google Scholar]
  24. Hassett D. J., Ma J. F., Elkins J. G., McDermott T. R., Ochsner U. A., West S. E., Huang C. T., Fredericks J., Burnett S.. other authors 1999; Quorum sensing in Pseudomonas aeruginosa controls expression of catalase and superoxide dismutase genes and mediates biofilm susceptibility to hydrogen peroxide. Mol Microbiol34:1082–1093[CrossRef]
    [Google Scholar]
  25. Hentzer M., Wu H., Andersen J. B., Riedel K., Rasmussen T. B., Bagge N., Kumar N., Schembri M. A., Song Z.. other authors 2003; Attenuation of Pseudomonas aeruginosa virulence by quorum sensing inhibitors. EMBO J22:3803–3815[CrossRef]
    [Google Scholar]
  26. Heydorn A., Nielsen A. T., Hentzer M., Sternberg C., Givskov M., Ersboll B. K., Molin S.. 2000; Quantification of biofilm structures by the novel computer program COMSTAT. Microbiology146:2395–2407
    [Google Scholar]
  27. Høiby N., Krogh Johansen H., Moser C., Song Z., Ciofu O., Kharazmi A.. 2001; Pseudomonas aeruginosa and the in vitro and in vivo biofilm mode of growth. Microbes Infect3:23–35[CrossRef]
    [Google Scholar]
  28. Johnson M., Cockayne A., Williams P. H., Morrissey J. A.. 2005; Iron-responsive regulation of biofilm formation in Staphylococcus aureus involves Fur-dependent and Fur-independent mechanisms. J Bacteriol187:8211–8215[CrossRef]
    [Google Scholar]
  29. Klausen M., Aaes-Jorgensen A., Molin S., Tolker-Nielsen T.. 2003a; Involvement of bacterial migration in the development of complex multicellular structures in Pseudomonas aeruginosa biofilms. Mol Microbiol50:61–68[CrossRef]
    [Google Scholar]
  30. Klausen M., Heydorn A., Ragas P., Lambertsen L., Aaes-Jorgensen A., Molin S., Tolker-Nielsen T.. 2003b; Biofilm formation by Pseudomonas aeruginosa wild type, flagella and type IV pili mutants. Mol Microbiol48:1511–1524[CrossRef]
    [Google Scholar]
  31. Lam J., Chan R., Lam K., Costerton J. W.. 1980; Production of mucoid microcolonies by Pseudomonas aeruginosa within infected lungs in cystic fibrosis. Infect Immun28:546–556
    [Google Scholar]
  32. Latifi A., Winson M. K., Foglino M., Bycroft B. W., Stewart G. S., Lazdunski A., Williams P.. 1995; Multiple homologues of LuxR and LuxI control expression of virulence determinants and secondary metabolites through quorum sensing in Pseudomonas aeruginosa PAO1. Mol Microbiol17:333–343[CrossRef]
    [Google Scholar]
  33. Latifi A., Foglino M., Tanaka K., Williams P., Lazdunski A.. 1996; A hierarchical quorum-sensing cascade in Pseudomonas aeruginosa links the transcriptional activators LasR and RhIR (VsmR) to expression of the stationary-phase sigma factor RpoS. Mol Microbiol21:1137–1146[CrossRef]
    [Google Scholar]
  34. Matsukawa M., Greenberg E. P.. 2004; Putative exopolysaccharide synthesis genes influence Pseudomonas aeruginosa biofilm development. J Bacteriol186:4449–4456[CrossRef]
    [Google Scholar]
  35. Miller J. H.. 1972; Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  36. Musk D. J., Banko D. A., Hergenrother P. J.. 2005; Iron salts perturb biofilm formation and disrupt existing biofilms of Pseudomonas aeruginosa. Chem Biol12:789–796[CrossRef]
    [Google Scholar]
  37. Nemoto K., Hirota K., Murakami K., Taniguti K., Murata H., Viducic D., Miyake Y.. 2003; Effect of Varidase (streptodornase) on biofilm formed by Pseudomonas aeruginosa. Chemotherapy49:121–125[CrossRef]
    [Google Scholar]
  38. Ochsner U. A., Wilderman P. J., Vasil A. I., Vasil M. L.. 2002; GeneChip expression analysis of the iron starvation response in Pseudomonas aeruginosa : identification of novel pyoverdine biosynthesis genes. Mol Microbiol45:1277–1287[CrossRef]
    [Google Scholar]
  39. O'Toole G. A., Kolter R.. 1998; Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol Microbiol30:295–304[CrossRef]
    [Google Scholar]
  40. Palmer K. L., Mashburn L. M., Singh P. K., Whiteley M.. 2005; Cystic fibrosis sputum supports growth and cues key aspects of Pseudomonas aeruginosa physiology. J Bacteriol187:5267–5277[CrossRef]
    [Google Scholar]
  41. 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. Science260:1127–1130[CrossRef]
    [Google Scholar]
  42. Pesci E. C., Milbank J. B., Pearson J. P., McKnight S., Kende A. S., Greenberg E. P., Iglewski B. H.. 1999; Quinolone signaling in the cell-to-cell communication system of Pseudomonas aeruginosa. Proc Natl Acad Sci U S A96:11229–11234[CrossRef]
    [Google Scholar]
  43. Ratledge C., Dover L. G.. 2000; Iron metabolism in pathogenic bacteria. Annu Rev Microbiol54:881–941[CrossRef]
    [Google Scholar]
  44. Rombel I. T., McMorran B. J., Lamont I. L.. 1995; Identification of a DNA sequence motif required for expression of iron-regulated genes in pseudomonads. Mol Gen Genet246:519–528[CrossRef]
    [Google Scholar]
  45. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: a Laboratory Manual , 2nd edn.vol. 1 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  46. Sauer K., Camper A. K., Ehrlich G. D., Costerton J. W., Davies D. G.. 2002; Pseudomonas aeruginosa displays multiple phenotypes during development as a biofilm. J Bacteriol184:1140–1154[CrossRef]
    [Google Scholar]
  47. Singh P. K., Parsek M. R., Greenberg E. P., Welsh M. J.. 2002; A component of innate immunity prevents bacterial biofilm development. Nature417:552–555[CrossRef]
    [Google Scholar]
  48. Sokol P. A., Cox C. D., Iglewski B. H.. 1982; Pseudomonas aeruginosa mutants altered in their sensitivity to the effect of iron on toxin A or elastase yields. J Bacteriol151:783–787
    [Google Scholar]
  49. Sriramulu D. D., Lam J. S., Lünsdorf H., Römling U.. 2005; Microcolony formation: a novel biofilm model of Pseudomonas aeruginosa for the cystic fibrosis lung. J Med Microbiol54:667–676[CrossRef]
    [Google Scholar]
  50. Sternberg C., Tolker-Nielsen T.. 2005; Growing and analyzing biofilms in flow cells. In Current Protocols in Microbiology1B.2.1–1B.2.15 Edited by Coico R., Kowalik T., Quarles J., Stevenson B., Taylor R.. New York: Wiley;
    [Google Scholar]
  51. Sutherland I. W.. 2001; The biofilm matrix – an immobilized but dynamic microbial environment. Arch Microbiol9:222–227
    [Google Scholar]
  52. Weinberg E. D.. 1999; Iron loading and disease surveillance. Emerg Infect Dis5:346–352[CrossRef]
    [Google Scholar]
  53. Whitchurch C. B., Tolker-Nielsen T., Ragas P. C., Mattick J. S.. 2002; Extracellular DNA required for bacterial biofilm formation. Science295:1487[CrossRef]
    [Google Scholar]
  54. Winson M. K., Camara M., Latifi A., Foglino M., Chhabra S. R., Daykin M., Bally M., Chapon V., Salmond G. P.. 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 U S A92:9427–9431[CrossRef]
    [Google Scholar]
  55. Worlitzsch D., Tarran R., Ulrich M., Schwab U., Cekici A., Meyer K. C., Birrer P., Bellon G., Berger J.. other authors 2002; Effects of reduced mucus oxygen concentration in airway Pseudomonas infections of cystic fibrosis patients. J Clin Invest109:317–325[CrossRef]
    [Google Scholar]
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