1887

Abstract

Uropathogenic (UPEC) causes more than 90 % of all human urinary tract infections through type 1 piliated UPEC cells binding to bladder epithelial cells. The FimB and FimE site-specific recombinases orient the element containing the structural gene promoter. Regulation of and depends on environmental pH and osmolality. The EnvZ/OmpR two-component system affects osmoregulation in . To ascertain if OmpR directly regulated the gene promoters, gel mobility shift and DNase I footprinting experiments were performed using OmpR or phosphorylated OmpR (OmpR-P) mixed with the promoter regions of UPEC strain NU149. Both OmpR-P and OmpR bound weakly to one promoter. Because there was weak binding to one promoter, strain NU149 was grown in different pH and osmolality environments, and total RNAs were extracted from each population and converted to cDNAs. Quantitative reverse-transcriptase PCR showed no differences in transcription among the different growth conditions. Conversely, Western blots showed a significant increase in OmpR protein in UPEC cells grown in a combined low pH/high osmolality environment versus a neutral pH/high osmolality environment. In a high osmolality environment, the mutant expressed more transcripts and Phase-ON positioning of the element as well as higher type 1 pili levels than wild-type cells. Together these results suggest that OmpR may be post-transcriptionally regulated in UPEC cells growing in a low pH/high osmolality environment, which regulates in UPEC.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.059386-0
2013-02-01
2024-04-27
Loading full text...

Full text loading...

/deliver/fulltext/micro/159/2/316.html?itemId=/content/journal/micro/10.1099/mic.0.059386-0&mimeType=html&fmt=ahah

References

  1. Abraham S. N., Babu J. P., Giampapa C. S., Hasty D. L., Simpson W. A., Beachey E. H. ( 1985a). Protection against Escherichia coli-induced urinary tract infections with hybridoma antibodies directed against type 1 fimbriae or complementary d-mannose receptors. Infect Immun 48:625–628[PubMed]
    [Google Scholar]
  2. Abraham J. M., Freitag C. S., Clements J. R., Eisenstein B. I. ( 1985b). An invertible element of DNA controls phase variation of type 1 fimbriae of Escherichia coli . Proc Natl Acad Sci U S A 82:5724–5727 [View Article][PubMed]
    [Google Scholar]
  3. Bang I. S., Kim B. H., Foster J. W., Park Y. K. ( 2000). OmpR regulates the stationary-phase acid tolerance response of Salmonella enterica serovar Typhimurium. J Bacteriol 182:2245–2252[PubMed] [CrossRef]
    [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[PubMed]
    [Google Scholar]
  5. Cai S. J., Inouye M. ( 2002). EnvZ–OmpR interaction and osmoregulation in Escherichia coli . J Biol Chem 277:24155–24161 [View Article][PubMed]
    [Google Scholar]
  6. Delgado J., Forst S., Harlocker S., Inouye M. ( 1993). Identification of a phosphorylation site and functional analysis of conserved aspartic acid residues of OmpR, a transcriptional activator for ompF and ompC in Escherichia coli . Mol Microbiol 10:1037–1047 [View Article][PubMed]
    [Google Scholar]
  7. 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[PubMed]
    [Google Scholar]
  8. El-Labany S., Sohanpal B. K., Lahooti M., Akerman R., Blomfield I. C. ( 2003). Distant cis-active sequences and sialic acid control the expression of fimB in Escherichia coli K-12. Mol Microbiol 49:1109–1118 [View Article][PubMed]
    [Google Scholar]
  9. Forst S., Delgado J., Ramakrishnan G., Inouye M. ( 1988). Regulation of ompC and ompF expression in Escherichia coli in the absence of envZ . J Bacteriol 170:5080–5085[PubMed]
    [Google Scholar]
  10. Forst S., Delgado J., Inouye M. ( 1989). Phosphorylation of OmpR by the osmosensor EnvZ modulates expression of the ompF and ompC genes in Escherichia coli . Proc Natl Acad Sci U S A 86:6052–6056 [View Article][PubMed]
    [Google Scholar]
  11. Foxman B. ( 2003). Epidemiology of urinary tract infections: incidence, morbidity, and economic costs. Dis Mon 49:53–70 [View Article][PubMed]
    [Google Scholar]
  12. Foxman B., Barlow R., D’Arcy H., Gillespie B., Sobel J. D. ( 2000). Urinary tract infection: self-reported incidence and associated costs. Ann Epidemiol 10:509–515 [View Article][PubMed]
    [Google Scholar]
  13. Freitag C. S., Abraham J. M., Clements J. R., Eisenstein B. I. ( 1985). Genetic analysis of the phase variation control of expression of type 1 fimbriae in Escherichia coli . J Bacteriol 162:668–675[PubMed]
    [Google Scholar]
  14. Guillier M., Gottesman S. ( 2006). Remodelling of the Escherichia coli outer membrane by two small regulatory RNAs. Mol Microbiol 59:231–247 [View Article][PubMed]
    [Google Scholar]
  15. Guillier M., Gottesman S. ( 2008). The 5′ end of two redundant sRNAs is involved in the regulation of multiple targets, including their own regulator. Nucleic Acids Res 36:6781–6794 [View Article][PubMed]
    [Google Scholar]
  16. Harlocker S. L., Bergstrom L., Inouye M. ( 1995). Tandem binding of six OmpR proteins to the ompF upstream regulatory sequence of Escherichia coli . J Biol Chem 270:26849–26856[PubMed] [CrossRef]
    [Google Scholar]
  17. Head C. G., Tardy A., Kenney L. J. ( 1998). Relative binding affinities of OmpR and OmpR-phosphate at the ompF and ompC regulatory sites. J Mol Biol 281:857–870 [View Article][PubMed]
    [Google Scholar]
  18. Hooton T. M., Stamm W. E. ( 1997). Diagnosis and treatment of uncomplicated urinary tract infection. Infect Dis Clin North Am 11:551–581 [View Article][PubMed]
    [Google Scholar]
  19. Huang K. J., Igo M. M. ( 1996). Identification of the bases in the ompF regulatory region, which interact with the transcription factor OmpR. J Mol Biol 262:615–628 [View Article][PubMed]
    [Google Scholar]
  20. Huang Y. J., Tsai T. Y., Pan T. M. ( 2007). Physiological response and protein expression under acid stress of Escherichia coli O157 : H7 TWC01 isolated from Taiwan. J Agric Food Chem 55:7182–7191 [View Article][PubMed]
    [Google Scholar]
  21. Hultgren S. J., Porter T. N., Schaeffer A. J., Duncan J. L. ( 1985). Role of type 1 pili and effects of phase variation on lower urinary tract infections produced by Escherichia coli . Infect Immun 50:370–377[PubMed]
    [Google Scholar]
  22. Igo M. M., Silhavy T. J. ( 1988). EnvZ, a transmembrane environmental sensor of Escherichia coli K-12, is phosphorylated in vitro . J Bacteriol 170:5971–5973[PubMed]
    [Google Scholar]
  23. Keith B. R., Maurer L., Spears P. A., Orndorff P. E. ( 1986). Receptor-binding function of type 1 pili effects bladder colonization by a clinical isolate of Escherichia coli . Infect Immun 53:693–696[PubMed]
    [Google Scholar]
  24. Kenney L. J., Bauer M. D., Silhavy T. J. ( 1995). Phosphorylation-dependent conformational changes in OmpR, an osmoregulatory DNA-binding protein of Escherichia coli . Proc Natl Acad Sci U S A 92:8866–8870 [View Article][PubMed]
    [Google Scholar]
  25. 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[PubMed]
    [Google Scholar]
  26. Kunin C. M., Chambers S. T. ( 1989). Osmoprotective properties for bacteria of renal papilla and urine: role of betaines as osmoprotectant molecules. Host–arasite Interactions in Urinary Tract Infections327–332 Kass E., Svanborg Eden C. Chicago: University of Chicago Press;
    [Google Scholar]
  27. Liu X., Ferenci T. ( 2001). An analysis of multifactorial influences on the transcriptional control of ompF and ompC porin expression under nutrient limitation. Microbiology 147:2981–2989[PubMed]
    [Google Scholar]
  28. Livak K. J., Schmittgen T. D. ( 2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods 25:402–408 [View Article][PubMed]
    [Google Scholar]
  29. Maeda S., Ozawa Y., Mizuno T., Mizushima S. ( 1988). Stereospecific positioning of the cis-acting sequence with respect to the canonical promoter is required for activation of the ompC gene by a positive regulator, OmpR, in Escherichia coli . J Mol Biol 202:433–441 [View Article][PubMed]
    [Google Scholar]
  30. Majdalani N., Cunning C., Sledjeski D., Elliott T., Gottesman S. ( 1998). DsrA RNA regulates translation of RpoS message by an anti-antisense mechanism, independent of its action as an antisilencer of transcription. Proc Natl Acad Sci U S A 95:12462–12467 [View Article][PubMed]
    [Google Scholar]
  31. Matsubara M., Mizuno T. ( 1999). EnvZ-independent phosphotransfer signaling pathway of the OmpR-mediated osmoregulatory expression of OmpC and OmpF in Escherichia coli . Biosci Biotechnol Biochem 63:408–414 [View Article][PubMed]
    [Google Scholar]
  32. Mattison K., Kenney L. J. ( 2002). Phosphorylation alters the interaction of the response regulator OmpR with its sensor kinase EnvZ. J Biol Chem 277:11143–11148 [View Article][PubMed]
    [Google Scholar]
  33. McClain M. S., Blomfield I. C., Eisenstein B. I. ( 1991). Roles of fimB and fimE in site-specific DNA inversion associated with phase variation of type 1 fimbriae in Escherichia coli . J Bacteriol 173:5308–5314[PubMed]
    [Google Scholar]
  34. McClain M. S., Blomfield I. C., Eberhardt K. J., Eisenstein B. I. ( 1993). Inversion-independent phase variation of type 1 fimbriae in Escherichia coli . J Bacteriol 175:4335–4344[PubMed]
    [Google Scholar]
  35. McCleary W. R., Stock J. B. ( 1994). Acetyl phosphate and the activation of two-component response regulators. J Biol Chem 269:31567–31572[PubMed]
    [Google Scholar]
  36. McVicker G., Sun L., Sohanpal B. K., Gashi K., Williamson R. A., Plumbridge J., Blomfield I. C. ( 2011). SlyA protein activates fimB gene expression and type 1 fimbriation in Escherichia coli K-12. J Biol Chem 286:32026–32035 [View Article][PubMed]
    [Google Scholar]
  37. Miller J. H. ( 1972). Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  38. Nogueira T., Springer M. ( 2000). Post-transcriptional control by global regulators of gene expression in bacteria. Curr Opin Microbiol 3:154–158 [View Article][PubMed]
    [Google Scholar]
  39. Ofek I., Beachey E. H. ( 1978). Mannose binding and epithelial cell adherence of Escherichia coli . Infect Immun 22:247–254[PubMed]
    [Google Scholar]
  40. 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 [View Article][PubMed]
    [Google Scholar]
  41. Oshima T., Aiba H., Masuda Y., Kanaya S., Sugiura M., Wanner B. L., Mori H., Mizuno T. ( 2002). Transcriptome analysis of all two-component regulatory system mutants of Escherichia coli K-12. Mol Microbiol 46:281–291 [View Article][PubMed]
    [Google Scholar]
  42. Pallesen L., Madsen O., Klemm P. ( 1989). Regulation of the phase switch controlling expression of type 1 fimbriae in Escherichia coli . Mol Microbiol 3:925–931 [View Article][PubMed]
    [Google Scholar]
  43. Perry A., Ofek I., Silverblatt F. J. ( 1983). Enhancement of mannose-mediated stimulation of human granulocytes by type 1 fimbriae aggregated with antibodies on Escherichia coli surfaces. Infect Immun 39:1334–1345[PubMed]
    [Google Scholar]
  44. Prohinar P., Forst S. A., Reed D., Mandic-Mulec I., Weiss J. ( 2002). OmpR-dependent and OmpR-independent responses of Escherichia coli to sublethal attack by the neutrophil bactericidal/permeability increasing protein. Mol Microbiol 43:1493–1504 [View Article][PubMed]
    [Google Scholar]
  45. Qin L., Yoshida T., Inouye M. ( 2001). The critical role of DNA in the equilibrium between OmpR and phosphorylated OmpR mediated by EnvZ in Escherichia coli . Proc Natl Acad Sci U S A 98:908–913[PubMed]
    [Google Scholar]
  46. Rampersaud A., Harlocker S. L., Inouye M. ( 1994). The OmpR protein of Escherichia coli binds to sites in the ompF promoter region in a hierarchical manner determined by its degree of phosphorylation. J Biol Chem 269:12559–12566[PubMed]
    [Google Scholar]
  47. Rhee J. E., Sheng W., Morgan L. K., Nolet R., Liao X., Kenney L. J. ( 2008). Amino acids important for DNA recognition by the response regulator OmpR. J Biol Chem 283:8664–8677 [View Article][PubMed]
    [Google Scholar]
  48. Ross D. L., Neely A. E. ( 1983). Renal function. Textbook of Urinalysis and Body Fluids97–112 Norwalk, VA: Appleton Century Crofts;
    [Google Scholar]
  49. Russo F. D., Silhavy T. J. ( 1991). EnvZ controls the concentration of phosphorylated OmpR to mediate osmoregulation of the porin genes. J Mol Biol 222:567–580 [View Article][PubMed]
    [Google Scholar]
  50. Schaeffer A. J., Amundsen S. K., Schmidt L. N. ( 1979). Adherence of Escherichia coli to human urinary tract epithelial cells. Infect Immun 24:753–759[PubMed]
    [Google Scholar]
  51. Schaeffer A. J., Schwan W. R., Hultgren S. J., Duncan J. L. ( 1987). Relationship of type 1 pilus expression in Escherichia coli to ascending urinary tract infections in mice. Infect Immun 55:373–380[PubMed]
    [Google Scholar]
  52. Schwan W. R. ( 2009). Survival of uropathogenic Escherichia coli in the murine urinary tract is dependent on OmpR. Microbiology 155:1832–1839 [View Article][PubMed]
    [Google Scholar]
  53. Schwan W. R. ( 2011). Regulation of fim genes in uropathogenic Escherichia coli . World J Clin Infect Dis 1:17–25 [View Article]
    [Google Scholar]
  54. Schwan W. R., Seifert H. S., Duncan J. L. ( 1992). Growth conditions mediate differential transcription of fim genes involved in phase variation of type 1 pili. J Bacteriol 174:2367–2375[PubMed]
    [Google Scholar]
  55. Schwan W. R., Seifert H. S., Duncan J. L. ( 1994). Analysis of the fimB promoter region involved in type 1 pilus phase variation in Escherichia coli . Mol Gen Genet 242:623–630 [View Article][PubMed]
    [Google Scholar]
  56. Schwan W. R., Lee J. L., Lenard F. A., Matthews B. T., Beck M. T. ( 2002). Osmolarity and pH growth conditions regulate fim gene transcription and type 1 pilus expression in uropathogenic Escherichia coli . Infect Immun 70:1391–1402 [View Article][PubMed]
    [Google Scholar]
  57. Schwan W. R., Shibata S., Aizawa S.-I., Wolfe A. J. ( 2007). The two-component response regulator RcsB regulates type 1 piliation in Escherichia coli . J Bacteriol 189:7159–7163 [View Article][PubMed]
    [Google Scholar]
  58. Shin S., Park C. ( 1995). Modulation of flagellar expression in Escherichia coli by acetyl phosphate and the osmoregulator OmpR. J Bacteriol 177:4696–4702[PubMed]
    [Google Scholar]
  59. Silverblatt F. J., Dreyer J. S., Schauer S. ( 1979). Effect of pili on susceptibility of Escherichia coli to phagocytosis. Infect Immun 24:218–223[PubMed]
    [Google Scholar]
  60. Stentebjerg-Olesen B., Chakraborty T., Klemm P. ( 2000). FimE-catalyzed off-to-on inversion of the type 1 fimbrial phase switch and insertion sequence recruitment in an Escherichia coli K-12 fimB strain. FEMS Microbiol Lett 182:319–325 [View Article][PubMed]
    [Google Scholar]
  61. Teng C. H., Cai M., Shin S., Xie Y., Kim K. J., Khan N. A., Di Cello F., Kim K. S. ( 2005). Escherichia coli K1 RS218 interacts with human brain microvascular endothelial cells via type 1 fimbria bacteria in the fimbriated state. Infect Immun 73:2923–2931 [View Article][PubMed]
    [Google Scholar]
  62. Tsung K., Brissette R. E., Inouye M. ( 1989). Identification of the DNA-binding domain of the OmpR protein required for transcriptional activation of the ompF and ompC genes of Escherichia coli by in vivo DNA footprinting. J Biol Chem 264:10104–10109[PubMed]
    [Google Scholar]
  63. Valentin-Hansen P., Johansen J., Rasmussen A. A. ( 2007). Small RNAs controlling outer membrane porins. Curr Opin Microbiol 10:152–155 [View Article][PubMed]
    [Google Scholar]
  64. van der Bosch J. F., Verboom-Sohmer U., Postma P., de Graaff J., MacLaren D. M. ( 1980). Mannose-sensitive and mannose-resistant adherence to human uroepithelial cells and urinary virulence of Escherichia coli . Infect Immun 29:226–233[PubMed]
    [Google Scholar]
  65. Virkola R., Westerlund B., Holthöfer H., Parkkinen J., Kekomäki M., Korhonen T. K. ( 1988). Binding characteristics of Escherichia coli adhesins in human urinary bladder. Infect Immun 56:2615–2622[PubMed]
    [Google Scholar]
  66. Walthers D., Carroll R. K., Navarre W. W., Libby S. J., Fang F. C., Kenney L. J. ( 2007). The response regulator SsrB activates expression of diverse Salmonella pathogenicity island 2 promoters and counters silencing by the nucleoid-associated protein H-NS. Mol Microbiol 65:477–493 [View Article][PubMed]
    [Google Scholar]
  67. Wolfe A. J. ( 2005). The acetate switch. Microbiol Mol Biol Rev 69:12–50 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.059386-0
Loading
/content/journal/micro/10.1099/mic.0.059386-0
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