1887

Abstract

In aquatic environments, biofilms constitute an ecological niche where persists as sessile cells. However, very little information on the sessile mode of life of is currently available. We report here the development of a model biofilm of strain Lens and the first transcriptome analysis of biofilm cells. Global gene expression analysis of sessile cells as compared to two distinct populations of planktonic cells revealed that a substantial proportion of genes is differentially expressed, as 2.3 % of the 2932 predicted genes exhibited at least a twofold change in gene expression. Comparison with previous results defining the gene expression profile of replicative- and transmissive-phase suggests that sessile cells resemble bacteria in the replicative phase. Further analysis of the most strongly regulated genes in sessile cells identified two induced gene clusters. One contains genes that encode alkyl hydroperoxide reductases known to act against oxidative stress. The second encodes proteins similar to PvcA and PvcB that are involved in siderophore biosynthesis in . Since iron has been reported to modify biofilm formation in other species, we further focused on iron control of gene expression and biofilm formation. Among the genes showing the greatest differences in expression between planktonic cells and biofilm, only and were regulated by iron concentration. A Δ mutant showed no changes in biofilm formation compared to the wild-type, suggesting that the product is not mandatory for biofilm formation. However, biofilm formation by wild-type and a Δ strain was clearly inhibited in iron-rich conditions.

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2008-01-01
2020-03-31
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References

  1. Abu Kwaik Y., Gao L. Y., Stone B. J., Venkataraman C., Harb O. S.. 1998; Invasion of protozoa by Legionella pneumophila and its role in bacterial ecology and pathogenesis. Appl Environ Microbiol64:3127–3133
    [Google Scholar]
  2. Allard K. A., Viswanathan V. K., Cianciotto N. P.. 2006; lbtA and lbtB are required for production of the Legionella pneumophila siderophore legiobactin. J Bacteriol188:1351–1363
    [Google Scholar]
  3. Andrews S. C., Robinson A. K., Rodriguez-Quinones F.. 2003; Bacterial iron homeostasis. FEMS Microbiol Rev27:215–237
    [Google Scholar]
  4. Baillon M. L., van Vliet A. H., Ketley J. M., Constantinidou C., Penn C. W.. 1999; An iron-regulated alkyl hydroperoxide reductase (AhpC) confers aerotolerance and oxidative stress resistance to the microaerophilic pathogen Campylobacter jejuni . J Bacteriol181:4798–4804
    [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
    [Google Scholar]
  6. Beloin C., Valle J., Latour-Lambert P., Faure P., Kzreminski M., Balestrino D., Haagensen J. A., Molin S., Prensier G.. other authors 2004; Global impact of mature biofilm lifestyle on Escherichia coli K-12 gene expression. Mol Microbiol51:659–674
    [Google Scholar]
  7. Brüggemann H., Hagman H., Jules M., Sismeiro O., Dillies M. A., Gouyette C., Kunst F., Steinert M., Heuner K.. other authors 2006; Virulence strategies for infecting phagocytes deduced from the in vivo transcriptional program of Legionella pneumophila . Cell Microbiol8:1228–1240
    [Google Scholar]
  8. Cazalet C., Rusniok C., Bruggemann H., Zidane N., Magnier A., Ma L., Tichit M., Jarraud S., Bouchier C.. other authors 2004; Evidence in the Legionella pneumophila genome for exploitation of host cell functions and high genome plasticity. Nat Genet36:1165–1173
    [Google Scholar]
  9. Chien M., Morozova I., Shi S., Sheng H., Chen J., Gomez S. M., Asamani G., Hill K., Nuara J.. other authors 2004; The genomic sequence of the accidental pathogen Legionella pneumophila . Science305:1966–1968
    [Google Scholar]
  10. Cianciotto N. P.. 2007; Iron acquisition by Legionella pneumophila . Biometals20:323–331
    [Google Scholar]
  11. De Buck E., Maes L., Meyen E., Van Mellaert L., Geukens N., Anne J., Lammertyn E.. 2005; Legionella pneumophila Philadelphia-1 tatB and tatC affect intracellular replication and biofilm formation. Biochem Biophys Res Commun331:1413–1420
    [Google Scholar]
  12. Declerck P., Behets J., van Hoef V., Ollevier F.. 2007; Detection of Legionella spp. and some of their amoeba hosts in floating biofilms from anthropogenic and natural aquatic environments. Water Res41:3159–3167
    [Google Scholar]
  13. Delmar P., Robin S., Daudin J. J.. 2005; VarMixt: efficient variance modelling for the differential analysis of replicated gene expression data. Bioinformatics21:502–508
    [Google Scholar]
  14. Domka J., Lee J., Bansal T., Wood T. K.. 2007; Temporal gene-expression in Escherichia coli K-12 biofilms. Environ Microbiol9:332–346
    [Google Scholar]
  15. Donlan R. M., Costerton J. W.. 2002; Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev15:167–193
    [Google Scholar]
  16. Fettes P. S., Forsbach-Birk V., Lynch D., Marre R.. 2001; Overexpresssion of a Legionella pneumophila homologue of the E. coli regulator csrA affects cell size, flagellation, and pigmentation. Int J Med Microbiol291:353–360
    [Google Scholar]
  17. Grifantini R., Frigimelica E., Delany I., Bartolini E., Giovinazzi S., Balloni S., Agarwal S., Galli G., Genco C.. other authors 2004; Characterization of a novel Neisseria meningitidis Fur and iron-regulated operon required for protection from oxidative stress: utility of DNA microarray in the assignment of the biological role of hypothetical genes. Mol Microbiol54:962–979
    [Google Scholar]
  18. Hall-Stoodley L., Costerton J. W., Stoodley P.. 2004; Bacterial biofilms: from the natural environment to infectious diseases. Nat Rev Microbiol2:95–108
    [Google Scholar]
  19. Heuner K., Brand B. C., Hacker J.. 1999; The expression of the flagellum of Legionella pneumophila is modulated by different environmental factors. FEMS Microbiol Lett175:69–77
    [Google Scholar]
  20. 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
    [Google Scholar]
  21. Kim B. R., Anderson J. E., Mueller S. A., Gaines W. A., Kendall A. M.. 2002; Literature review – efficacy of various disinfectants against Legionella in water systems. Water Res36:4433–4444
    [Google Scholar]
  22. Kuiper M. W., Wullings B. A., Akkermans A. D., Beumer R. R., van der Kooij D.. 2004; Intracellular proliferation of Legionella pneumophila in Hartmannella vermiformis in aquatic biofilms grown on plasticized polyvinyl chloride. Appl Environ Microbiol70:6826–6833
    [Google Scholar]
  23. LeBlanc J. J., Davidson R. J., Hoffman P. S.. 2006; Compensatory functions of two alkyl hydroperoxide reductases in the oxidative defense system of Legionella pneumophila . J Bacteriol188:6235–6244
    [Google Scholar]
  24. Livak K. J., Schmittgen T. D.. 2001; Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods25:402–408
    [Google Scholar]
  25. Mampel J., Spirig T., Weber S. S., Haagensen J. A., Molin S., Hilbi H.. 2006; Planktonic replication is essential for biofilm formation by Legionella pneumophila in a complex medium under static and dynamic flow conditions. Appl Environ Microbiol72:2885–2895
    [Google Scholar]
  26. Milohanic E., Glaser P., Coppee J. Y., Frangeul L., Vega Y., Vazquez-Boland J. A., Kunst F., Cossart P., Buchrieser C.. 2003; Transcriptome analysis of Listeria monocytogenes identifies three groups of genes differently regulated by PrfA. Mol Microbiol47:1613–1625
    [Google Scholar]
  27. Molofsky A. B., Swanson M. S.. 2003; Legionella pneumophila CsrA is a pivotal repressor of transmission traits and activator of replication. Mol Microbiol50:445–461
    [Google Scholar]
  28. Molofsky A. B., Swanson M. S.. 2004; Differentiate to thrive: lessons from the Legionella pneumophila life cycle. Mol Microbiol53:29–40
    [Google Scholar]
  29. Murga R., Forster T. S., Brown E., Pruckler J. M., Fields B. S., Donlan R. M.. 2001; Role of biofilms in the survival of Legionella pneumophila in a model potable-water system. Microbiology147:3121–3126
    [Google Scholar]
  30. 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
    [Google Scholar]
  31. O'Toole G. A., Kolter R.. 1998; Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol Microbiol30:295–304
    [Google Scholar]
  32. Pham T. H., Webb J. S., Rehm B. H.. 2004; The role of polyhydroxyalkanoate biosynthesis by Pseudomonas aeruginosa in rhamnolipid and alginate production as well as stress tolerance and biofilm formation. Microbiology150:3405–3413
    [Google Scholar]
  33. Piao Z., Sze C. C., Barysheva O., Iida K., Yoshida S.. 2006; Temperature-regulated formation of mycelial mat-like biofilms by Legionella pneumophila . Appl Environ Microbiol72:1613–1622
    [Google Scholar]
  34. Pratt L. A., Kolter R.. 1998; Genetic analysis of Escherichia coli biofilm formation: roles of flagella, motility, chemotaxis and type I pili. Mol Microbiol30:285–293
    [Google Scholar]
  35. Reiner A., Yekutieli D., Benjamini Y.. 2003; Identifying differentially expressed genes using false discovery rate controlling procedures. Bioinformatics19:368–375
    [Google Scholar]
  36. Robey M., Cianciotto N. P.. 2002; Legionella pneumophila feoAB promotes ferrous iron uptake and intracellular infection. Infect Immun70:5659–5669
    [Google Scholar]
  37. Rocha E. R., Smith C. J.. 1999; Role of the alkyl hydroperoxide reductase ( ahpCF ) gene in oxidative stress defense of the obligate anaerobe Bacteroides fragilis . J Bacteriol181:5701–5710
    [Google Scholar]
  38. Saby S., Vidal A., Suty H.. 2005; Resistance of Legionella to disinfection in hot water distribution systems. Water Sci Technol52:15–28
    [Google Scholar]
  39. 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
    [Google Scholar]
  40. Sauer K., Cullen M. C., Rickard A. H., Zeef L. A., Davies D. G., Gilbert P.. 2004; Characterization of nutrient-induced dispersion in Pseudomonas aeruginosa PAO1 biofilm. J Bacteriol186:7312–7326
    [Google Scholar]
  41. Schembri M. A., Kjaergaard K., Klemm P.. 2003; Global gene expression in Escherichia coli biofilms. Mol Microbiol48:253–267
    [Google Scholar]
  42. Shirtliff M. E., Mader J. T., Camper A. K.. 2002; Molecular interactions in biofilms. Chem Biol9:859–871
    [Google Scholar]
  43. Singh P. K.. 2004; Iron sequestration by human lactoferrin stimulates P. aeruginosa surface motility and blocks biofilm formation. Biometals17:267–270
    [Google Scholar]
  44. Soderberg M. A., Rossier O., Cianciotto N. P.. 2004; The type II protein secretion system of Legionella pneumophila promotes growth at low temperatures. J Bacteriol186:3712–3720
    [Google Scholar]
  45. Steinert M., Hentschel U., Hacker J.. 2002; Legionella pneumophila : an aquatic microbe goes astray. FEMS Microbiol Rev26:149–162
    [Google Scholar]
  46. Stintzi A., Cornelis P., Hohnadel D., Meyer J. M., Dean C., Poole K., Kourambas S., Krishnapillai V.. 1996; Novel pyoverdine biosynthesis gene(s) of Pseudomonas aeruginosa PAO. Microbiology142:1181–1190
    [Google Scholar]
  47. Stintzi A., Johnson Z., Stonehouse M., Ochsner U., Meyer J. M., Vasil M. L., Poole K.. 1999; The pvc gene cluster of Pseudomonas aeruginosa : role in synthesis of the pyoverdine chromophore and regulation by PtxR and PvdS. J Bacteriol181:4118–4124
    [Google Scholar]
  48. Thomas V., Bouchez T., Nicolas V., Robert S., Loret J. F., Levi Y.. 2004; Amoebae in domestic water systems: resistance to disinfection treatments and implication in Legionella persistence. J Appl Microbiol97:950–963
    [Google Scholar]
  49. Visca P., Ciervo A., Orsi N.. 1994; Cloning and nucleotide sequence of the pvdA gene encoding the pyoverdin biosynthetic enzyme l-ornithine N 5-oxygenase in Pseudomonas aeruginosa . J Bacteriol176:1128–1140
    [Google Scholar]
  50. Waite R. D., Papakonstantinopoulou A., Littler E., Curtis M. A.. 2005; Transcriptome analysis of Pseudomonas aeruginosa growth: comparison of gene expression in planktonic cultures and developing and mature biofilms. J Bacteriol187:6571–6576
    [Google Scholar]
  51. Watnick P. I., Kolter R.. 1999; Steps in the development of a Vibrio cholerae El Tor biofilm. Mol Microbiol34:586–595
    [Google Scholar]
  52. Watnick P., Kolter R.. 2000; Biofilm, city of microbes. J Bacteriol182:2675–2679
    [Google Scholar]
  53. Whiteley M., Bangera M. G., Bumgarner R. E., Parsek M. R., Teitzel G. M., Lory S., Greenberg E. P.. 2001; Gene expression in Pseudomonas aeruginosa biofilms. Nature413:860–864
    [Google Scholar]
  54. Yang Y. H., Dudoit S., Luu P., Lin D. M., Peng V., Ngai J., Speed T. P.. 2002; Normalization for cDNA microarray data: a robust composite method addressing single and multiple slide systematic variation. Nucleic Acids Res30:e15
    [Google Scholar]
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