1887

Abstract

Type 1 fimbriae of facilitate attachment to the host mucosa and promote biofilm formation on abiotic surfaces. The transcriptional regulator LrhA, which is known as a repressor of flagellar, motility and chemotaxis genes, regulates biofilm formation and expression of type 1 fimbriae. Whole-genome expression profiling revealed that inactivation of results in an increased expression of structural components of type 1 fimbriae. , LrhA bound to the promoter regions of the two recombinases (FimB and FimE) that catalyse the inversion of the promoter, and to the invertible element itself. Translational fusions with these genes and quantification of transcript levels by real-time PCR showed that LrhA influences type 1 fimbrial phase variation, primarily via activation of FimE, which is required for the ON-to-OFF transition of the switch. Enhanced type 1 fimbrial expression as a result of disruption was confirmed by mannose-sensitive agglutination of yeast cells. Biofilm formation was stimulated by inactivation and completely suppressed upon LrhA overproduction. The effects of LrhA on biofilm formation were exerted via the changed levels of surface molecules, most probably both flagella and type 1 fimbriae. Together, the data show a role for LrhA as a repressor of type 1 fimbrial expression, and thus as a regulator of the initial stages of biofilm development and, presumably, bacterial adherence to epithelial host cells also.

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2005-10-01
2019-09-23
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References

  1. Allison, C., Emödy, L., Coleman, N. & Hughes, C. ( 1994; ). The role of swarm cell differentiation and multicellular migration in the uropathogenicity of Proteus mirabilis. J Infect Dis 169, 1155–1158.[CrossRef]
    [Google Scholar]
  2. Bahrani-Mougeot, F. K., Buckles, E. L., Lockatell, C. V., Hebel, J. R., Johnson, D. E., Tang, C. M. & Donnenberg, M. S. ( 2002; ). Type 1 fimbriae and extracellular polysaccharides are preeminent uropathogenic Escherichia coli virulence determinants in the murine urinary tract. Mol Microbiol 45, 1079–1093.[CrossRef]
    [Google Scholar]
  3. Berger, H., Hacker, J., Juarez, A., Hughes, C. & Goebel, W. ( 1982; ). Cloning of chromosomal determinants encoding hemolysin production and mannose-resistant hemagglutination in Escherichia coli. J Bacteriol 152, 1241–1247.
    [Google Scholar]
  4. Blomfield, I. C., McClain, M. S., Princ, J. A., Calie, P. J. & Eisenstein, B. I. ( 1991; ). Type 1 fimbriation and fimE mutants of Escherichia coli K-12. J Bacteriol 173, 5298–5307.
    [Google Scholar]
  5. Blomfield, I. C., Calie, P. J., Eberhardt, K. J., McClain, M. S. & Eisenstein, B. I. ( 1993; ). Lrp stimulates phase variation of type 1 fimbriation in Escherichia coli K-12. J Bacteriol 175, 27–36.
    [Google Scholar]
  6. Blomfield, I. C., Kulasekara, D. H. & Eisenstein, B. I. ( 1997; ). Integration host factor stimulates both FimB- and FimE-mediated site-specific DNA inversion that controls phase variation of type 1 fimbriae expression in Escherichia coli. Mol Microbiol 23, 705–717.[CrossRef]
    [Google Scholar]
  7. Bongaerts, J., Zoske, S., Weidner, U. & Unden, G. ( 1995; ). Transcriptional regulation of the proton translocating NADH dehydrogenase genes (nuoA-N) of Escherichia coli by electron acceptors, electron donors and gene regulators. Mol Microbiol 16, 521–534.[CrossRef]
    [Google Scholar]
  8. Brinton, C. C. ( 1959; ). Non flagellar appendages of bacteria. Nature 183, 782–786.[CrossRef]
    [Google Scholar]
  9. Connell, I., Agace, W., Klemm, P., Schembri, M., Marild, S. & Svanborg, C. ( 1996; ). Type 1 fimbrial expression enhances Escherichia coli virulence for the urinary tract. Proc Natl Acad Sci U S A 93, 9827–9832.[CrossRef]
    [Google Scholar]
  10. Dobrindt, U., Blum-Oehler, G., Nagy, G., Schneider, G., Johann, A., Gottschalk, G. & Hacker, J. ( 2002; ). Genetic structure and distribution of four pathogenicity islands (PAI I536 to PAI IV536) of uropathogenic Escherichia coli strain 536. Infect Immun 70, 6365–6372.[CrossRef]
    [Google Scholar]
  11. Dobrindt, U., Agerer, F., Michaelis, K. & 7 other authors ( 2003; ). Analysis of genome plasticity in pathogenic and commensal Escherichia coli isolates by use of DNA arrays. J Bacteriol 185, 1831–1840.[CrossRef]
    [Google Scholar]
  12. Donato, G. M., Lelivelt, M. J. & Kawula, T. H. ( 1997; ). Promoter-specific repression of fimB expression by the Escherichia coli nucleoid-associated protein H-NS. J Bacteriol 179, 6618–6625.
    [Google Scholar]
  13. Donnenberg, M. S. & Kaper, J. B. ( 1991; ). Construction of an eae deletion mutant of enteropathogenic Escherichia coli by using a positive-selection suicide vector. Infect Immun 59, 4310–4317.
    [Google Scholar]
  14. Dorman, C. J. & Higgins, C. F. ( 1987; ). Fimbrial phase variation in Escherichia coli: dependence of integration host factor and homologies with other site-specific recombinases. J Bacteriol 169, 3840–3843.
    [Google Scholar]
  15. Drapal, N. & Sawers, G. ( 1995; ). Promoter 7 of the Escherichia coli pfl operon is a major determinant in the anaerobic regulation of expression by ArcA. J Bacteriol 177, 5338–5341.
    [Google Scholar]
  16. Gally, D. L., Rucker, T. J. & Blomfield, I. C. ( 1994; ). The leucine-responsive regulatory protein binds to the fim switch to control phase variation of type 1 fimbrial expression in Escherichia coli K-12. J Bacteriol 176, 5665–5672.
    [Google Scholar]
  17. Gally, D. L., Leathart, J. & Blomfield, I. C. ( 1996; ). Interaction of FimB and FimE with the fim switch that controls the phase variation of type 1 fimbriae in Escherichia coli K-12. Mol Microbiol 21, 725–738.[CrossRef]
    [Google Scholar]
  18. Gibson, K. E. & Silhavy, T. J. ( 1999; ). The LysR homolog LrhA promotes RpoS degradation by modulating activity of the response regulator sprE. J Bacteriol 181, 563–571.
    [Google Scholar]
  19. Gunther, N. W., 4th, Lockatell, V., Johnson, D. E. & Mobley, H. L. T. ( 2001; ). In vivo dynamics of type 1 fimbria regulation in uropathogenic Escherichia coli during experimental urinary tract infection. Infect Immun 69, 2838–2846.[CrossRef]
    [Google Scholar]
  20. Harris, S. J., Shih, Y., Bentley, S. D. & Salmond, P. P. C. ( 1998; ). The hexA gene of Erwinia carotovora encodes a LysR homologue and regulates motility and the expression of multiple virulence determinants. Mol Microbiol 28, 705–717.
    [Google Scholar]
  21. Jackson, D. W., Suzuki, K., Oakford, L., Simecka, J. W., Hart, M. E. & Romeo, T. ( 2002; ). Biofilm formation and dispersal under the influence of the global regulator CsrA of Escherichia coli. J Bacteriol 184, 290–301.[CrossRef]
    [Google Scholar]
  22. Jensen, K. F. ( 1993; ). The Escherichia coli K-12 ‘wild-types' W3110 and MG1655 have an rph frameshift mutation that leads to pyrimidine starvation due to low pyrE expression levels. J Bacteriol 175, 3401–3407.
    [Google Scholar]
  23. Jones, C. H., Pinkner, J. S., Roth, R., Heuser, J., Nicholes, A. V., Abraham, S. N. & Hultgren, S. J. ( 1995; ). FimH adhesin of type 1 pili is assembled into a fibrillar tip structure in the Enterobacteriaceae. Proc Natl Acad Sci U S A 92, 2081–2085.[CrossRef]
    [Google Scholar]
  24. Kalir, S., McClure, J., Pabbaraju, K., Southward, C., Ronen, M., Leibler, S., Surette, M. G. & Alon, U. ( 2001; ). Ordering genes in a flagella pathway by analysis of expression kinetics from living bacteria. Science 292, 2080–2083.[CrossRef]
    [Google Scholar]
  25. Keith, B. R., Maurer, L., Spears, P. A. & Orndorff, P. E. ( 1986; ). Receptor-binding function of type 1 pili affects bladder colonization by a clinical isolate of Escherichia coli. Infect Immun 53, 693–696.
    [Google Scholar]
  26. Klemm, P. ( 1986; ). Two regulatory fim genes, fimB and fimE, control the phase variation of type 1 fimbriae in Escherichia coli. EMBO J 5, 1389–1393.
    [Google Scholar]
  27. Lehnen, D., Blumer, C., Polen, T., Wackwitz, B., Wendisch, V. F. & Unden, G. ( 2002; ). LrhA as a new transcriptional key regulator of flagella, motility and chemotaxis genes in Escherichia coli. Mol Microbiol 45, 521–532.[CrossRef]
    [Google Scholar]
  28. Macnab, R. M. ( 1996; ). Flagella and motility. In Escherichia coli and Salmonella, pp. 123–145. Edited by F. C. Neidhardt. Washington, DC: American Society for Microbiology.
  29. Martinez, J. J., Mulvey, M. A., Schilling, J. D., Pinkner, J. S. & Hultgren, S. J. ( 2000; ). Type 1 pilus-mediated bacterial invasion of bladder epithelial cells. EMBO J 19, 2803–2812.[CrossRef]
    [Google Scholar]
  30. Miller, J. H. ( 1992; ). A Short Course in Bacterial Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  31. Miller, V. L. & Mekalanos, J. J. ( 1988; ). A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR. J Bacteriol 170, 2575–2583.
    [Google Scholar]
  32. Mukherjee, A., Cui, Y., Ma, W., Liu, Y. & Chatterjee, A. K. ( 2000; ). hexA of Erwinia carotovora ssp. carotovora strain Ecc71 negatively regulates production of RpoS and rsmB RNA, a global regulator of extracellular proteins, plant virulence and the quorum-sensing signal, N-(3-oxohexanoyl)-l-homoserine lactone. Environ Microbiol 2, 203–215.[CrossRef]
    [Google Scholar]
  33. Mulvey, M. A. ( 2002; ). Adhesion and entry of uropathogenic Escherichia coli. Cell Microbiol 4, 257–271.[CrossRef]
    [Google Scholar]
  34. Mulvey, M. A., Lopez-Boado, Y. S., Wilson, C. L., Roth, R., Parks, W. C., Heuser, J. & Hultgren, S. J. ( 1998; ). Induction and evasion of host defenses by type 1-piliated uropathogenic Escherichia coli. Science 282, 1494–1497.[CrossRef]
    [Google Scholar]
  35. Nagy, G., Dobrindt, U., Schneider, G., Khan, A. S., Hacker, J. & Emödy, L. ( 2002; ). Loss of regulatory protein RfaH attenuates virulence of uropathogenic Escherichia coli. Infect Immun 70, 4406–4413.[CrossRef]
    [Google Scholar]
  36. O'Gara, J. P. & Dorman, C. J. ( 2000; ). Effects of local transcription and H-NS on inversion of the fim switch of Escherichia coli. Mol Microbiol 36, 457–466.[CrossRef]
    [Google Scholar]
  37. Olsen, P. B. & Klemm, P. ( 1994; ). Localization of promoters in the fim gene cluster and the effect of H-NS on the transcription of fimB and fimE. FEMS Microbiol Lett 116, 95–100.[CrossRef]
    [Google Scholar]
  38. Ostrow, K. S., Silhavy, T. J. & Garrett, S. ( 1986; ). cis-acting sites required for osmoregulation of ompF expression in Escherichia coli K-12. J Bacteriol 168, 1165–1171.
    [Google Scholar]
  39. O'Toole, G. A. & Kolter, R. ( 1998; ). Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis. Mol Microbiol 28, 449–461.[CrossRef]
    [Google Scholar]
  40. Pratt, L. A. & Kolter, R. ( 1998; ). Genetic analysis of Escherichia coli biofilm formation: roles of flagella, motility, chemotaxis and type 1 pili. Mol Microbiol 30, 285–293.[CrossRef]
    [Google Scholar]
  41. Sambrook, J., Fritsch, E. F. & Maniatis, T. ( 1989; ). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  42. Scharf, B., Schuster-Wolff-Buhring, H., Rachel, R. & Schmitt, R. ( 2001; ). Mutational analysis of the Rhizobium lupini H13-3 and Sinorhizobium meliloti flagellin genes: importance of flagellin A for flagellar filament structure and transcriptional regulation. J Bacteriol 183, 5334–5342.[CrossRef]
    [Google Scholar]
  43. Schembri, M. A. & Klemm, P. ( 2001; ). Biofilm formation in a hydrodynamic environment by novel fimH variants and ramifications for virulence. Infect Immun 69, 1322–1328.[CrossRef]
    [Google Scholar]
  44. Silhavy, T. J., Berman, M. & Enquist, L. W. ( 1984; ). Experiments with Gene Fusions. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  45. Simon, R., Priefer, U. & Pühler, A. ( 1983; ). A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram-negative bacteria. Bio/Technology 1, 781–791.
    [Google Scholar]
  46. Six, S., Andrews, S. C., Unden, G. & Guest, J. R. ( 1994; ). Escherichia coli possesses two homologous anaerobic C4-dicarboxylate membrane transporters (DcuA and DcuB) distinct from the aerobic dicarboxylate transport system (Dct). J Bacteriol 176, 6470–6478.
    [Google Scholar]
  47. Sohanpal, B. K., Kulasekara, H. D., Bonnen, A. & Blomfield, I. C. ( 2001; ). Orientational control of fimE expression in Escherichia coli. Mol Microbiol 42, 483–494.[CrossRef]
    [Google Scholar]
  48. Surgey, N., Robert-Baudouy, J. & Condemine, G. ( 1996; ). The Erwinia chrysanthemi pecT gene regulates pectinase gene expression. J Bacteriol 178, 1593–1599.
    [Google Scholar]
  49. Tusher, V. G., Tibshirani, R. & Chu, G. ( 2001; ). Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci U S A 98, 5116–5121.[CrossRef]
    [Google Scholar]
  50. Vidal, O., Longin, R., Prigent-Combaret, C., Dorel, C., Hooreman, M. & Lejeune, P. ( 1998; ). Isolation of an Escherichia coli K-12 mutant strain able to form biofilms on inert surfaces: involvement of a new ompR allele that increases curli expression. J Bacteriol 180, 2442–2449.
    [Google Scholar]
  51. Wendisch, V. F., Zimmer, D. P., Khodursky, A. B., Peter, B. J., Cozzarelli, N. & Kustu, S. ( 2001; ). Isolation of Escherichia coli mRNA and comparison of expression using mRNA and total RNA on DNA microarrays. Anal Biochem 290, 205–213.[CrossRef]
    [Google Scholar]
  52. Xia, Y., Gally, D., Forsman-Semb, K. & Uhlin, B. E. ( 2000; ). Regulatory cross-talk between adhesion operons in Escherichia coli: inhibition of type 1 fimbriae expression by the PapB protein. EMBO J 19, 1450–1457.[CrossRef]
    [Google Scholar]
  53. Yanisch-Perron, C., Vieira, J. & Messing, J. ( 1985; ). Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33, 103–119.[CrossRef]
    [Google Scholar]
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