1887

Abstract

is a pathogen of rodents, which causes diverse enteric and hepatic inflammatory diseases and malignancies. The urease enzyme is an important colonization factor of gastric species like , but little is known about the role and regulation of urease in enterohepatic species. Here it is reported that urease activity of does not contribute to acid resistance, and that it is nickel-responsive at the post-translational level. strain ATCC 51449 did not grow or survive at pH 3·0, and supplementation with urea or NiCl did not abrogate this acid sensitivity. Furthermore, urease enzyme activity of was acid-independent, which contrasts with the acid-induced urease system of . Nickel supplementation of Brucella medium resulted in a tenfold increase in urease activity in both and , but the maximum level of urease activity in was still three- to fivefold lower when compared to in the same conditions. The increase in urease activity of was not associated with elevation of urease mRNA or protein levels. Inhibition of protein synthesis by chloramphenicol did not affect nickel-responsive induction of urease activity in , and confirmed that nickel induction occurs at the post-translational level, probably by activation of preformed apo-enzyme. In conclusion, both the role of the urease enzyme and the regulation of urease activity differ between the enterohepatic pathogen and the gastric pathogen .

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.28188-0
2005-12-01
2019-09-16
Loading full text...

Full text loading...

/deliver/fulltext/micro/151/12/3989.html?itemId=/content/journal/micro/10.1099/mic.0.28188-0&mimeType=html&fmt=ahah

References

  1. Akada, J. K., Shirai, M., Takeuchi, H., Tsuda, M. & Nakazawa, T. ( 2000; ). Identification of the urease operon in Helicobacter pylori and its control by mRNA decay in response to pH. Mol Microbiol 36, 1071–1084.[CrossRef]
    [Google Scholar]
  2. Andrutis, K. A., Fox, J. G., Schauer, D. B., Marini, R. P., Murphy, J. C., Yan, L. & Solnick, J. V. ( 1995; ). Inability of an isogenic urease-negative mutant stain of Helicobacter mustelae to colonize the ferret stomach. Infect Immun 63, 3722–3725.
    [Google Scholar]
  3. Baillon, M. L. A., van Vliet, A. H. M., 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 Bacteriol 181, 4798–4804.
    [Google Scholar]
  4. Beckwith, C. S., McGee, D. J., Mobley, H. L. & Riley, L. K. ( 2001; ). Cloning, expression, and catalytic activity of Helicobacter hepaticus urease. Infect Immun 69, 5914–5920.[CrossRef]
    [Google Scholar]
  5. Blaser, M. J. & Atherton, J. C. ( 2004; ). Helicobacter pylori persistence: biology and disease. J Clin Invest 113, 321–333.[CrossRef]
    [Google Scholar]
  6. Burne, R. A. & Chen, Y. Y. ( 2000; ). Bacterial ureases in infectious diseases. Microbes Infect 2, 533–542.[CrossRef]
    [Google Scholar]
  7. Bury-Mone, S., Skouloubris, S., Dauga, C., Thiberge, J. M., Dailidiene, D., Berg, D. E., Labigne, A. & De Reuse, H. ( 2003; ). Presence of active aliphatic amidases in Helicobacter species able to colonize the stomach. Infect Immun 71, 5613–5622.[CrossRef]
    [Google Scholar]
  8. Bury-Mone, S., Thiberge, J. M., Contreras, M., Maitournam, A., Labigne, A. & De Reuse, H. ( 2004; ). Responsiveness to acidity via metal ion regulators mediates virulence in the gastric pathogen Helicobacter pylori. Mol Microbiol 53, 623–638.[CrossRef]
    [Google Scholar]
  9. Chen, Y. Y. & Burne, R. A. ( 2003; ). Identification and characterization of the nickel uptake system for urease biogenesis in Streptococcus salivarius 57.I. J Bacteriol 185, 6773–6779.[CrossRef]
    [Google Scholar]
  10. Ernst, F. D., Kuipers, E. J., Heijens, A., Sarwari, R., Stoof, J., Penn, C. W., Kusters, J. G. & van Vliet, A. H. M. ( 2005; ). The nickel-responsive regulator NikR controls activation and repression of gene transcription in Helicobacter pylori. Infect Immun 73, 7252–7258.[CrossRef]
    [Google Scholar]
  11. Fox, J. G., Dewhirst, F. E., Tully, J. G., Paster, B. J., Yan, L., Taylor, N. S., Collins, M. J., Jr, Gorelick, P. L. & Ward, J. M. ( 1994; ). Helicobacter hepaticus sp. nov., a microaerophilic bacterium isolated from livers and intestinal mucosal scrapings from mice. J Clin Microbiol 32, 1238–1245.
    [Google Scholar]
  12. Franklin, C. L., Riley, L. K., Livingston, R. S., Beckwith, C. S., Besch-Williford, C. L. & Hook, R. R., Jr (1998; ). Enterohepatic lesions in SCID mice infected with Helicobacter bilis. Lab Anim Sci 48, 334–339.
    [Google Scholar]
  13. Franklin, C. L., Riley, L. K., Livingston, R. S., Beckwith, C. S., Hook, R. R., Jr, Besch-Williford, C. L., Hunziker, R. & Gorelick, P. L. ( 1999; ). Enteric lesions in SCID mice infected with “Helicobacter typhlonicus”, a novel urease-negative Helicobacter species. Lab Anim Sci 49, 496–505.
    [Google Scholar]
  14. Kusters, J. G., Gerrits, M. M., Van Strijp, J. A. & Vandenbroucke-Grauls, C. M. J. E. ( 1997; ). Coccoid forms of Helicobacter pylori are the morphologic manifestation of cell death. Infect Immun 65, 3672–3679.
    [Google Scholar]
  15. Kuwahara, H., Miyamoto, Y., Akaike, T., Kubota, T., Sawa, T., Okamoto, S. & Maeda, H. ( 2000; ). Helicobacter pylori urease suppresses bactericidal activity of peroxynitrite via carbon dioxide production. Infect Immun 68, 4378–4383.[CrossRef]
    [Google Scholar]
  16. Lee, A., Chen, M., Coltro, N., O'Rourke, J., Hazell, S., Hu, P. & Li, Y. ( 1993; ). Long term infection of the gastric mucosa with Helicobacter species does induce atrophic gastritis in an animal model of Helicobacter pylori infection. Zentralbl Bakteriol 280, 38–50.[CrossRef]
    [Google Scholar]
  17. Marcus, E. A., Moshfegh, A. P., Sachs, G. & Scott, D. R. ( 2005; ). The periplasmic alpha-carbonic anhydrase activity of Helicobacter pylori is essential for acid acclimation. J Bacteriol 187, 729–738.[CrossRef]
    [Google Scholar]
  18. Maurer, K. J., Ihrig, M. M., Rogers, A. B., Ng, V., Bouchard, G., Leonard, M. R., Carey, M. C. & Fox, J. G. ( 2005; ). Identification of cholelithogenic enterohepatic Helicobacter species and their role in murine cholesterol gallstone formation. Gastroenterology 128, 1023–1033.[CrossRef]
    [Google Scholar]
  19. Mehta, N. S., Benoit, S., Mysore, J. V., Sousa, R. S. & Maier, R. J. ( 2005; ). Helicobacter hepaticus hydrogenase mutants are deficient in hydrogen-supported amino acid uptake and in causing liver lesions in A/J mice. Infect Immun 73, 5311–5318.[CrossRef]
    [Google Scholar]
  20. Nilsson, H. O., Castedal, M., Olsson, R. & Wadstrom, T. ( 1999; ). Detection of Helicobacter in the liver of patients with chronic cholestatic liver diseases. J Physiol Pharmacol 50, 875–882.
    [Google Scholar]
  21. Queiroz, D. M. & Santos, A. ( 2001; ). Isolation of a Helicobacter strain from the human liver. Gastroenterology 121, 1023–1024.
    [Google Scholar]
  22. Sambrook, J., Fritsch, E. F. & Maniatis, T. ( 1989; ). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  23. Scott, D. R., Marcus, E. A., Weeks, D. L., Lee, A., Melchers, K. & Sachs, G. ( 2000; ). Expression of the Helicobacter pylori ureI gene is required for acidic pH activation of cytoplasmic urease. Infect Immun 68, 470–477.[CrossRef]
    [Google Scholar]
  24. Scott, D. R., Marcus, E. A., Weeks, D. L. & Sachs, G. ( 2002; ). Mechanisms of acid resistance due to the urease system of Helicobacter pylori. Gastroenterology 123, 187–195.[CrossRef]
    [Google Scholar]
  25. Skouloubris, S., Thiberge, J. M., Labigne, A. & De Reuse, H. ( 1998; ). The Helicobacter pylori UreI protein is not involved in urease activity but is essential for bacterial survival in vivo. Infect Immun 66, 4517–4521.
    [Google Scholar]
  26. Solnick, J. V. & Schauer, D. B. ( 2001; ). Emergence of diverse Helicobacter species in the pathogenesis of gastric and enterohepatic diseases. Clin Microbiol Rev 14, 59–97.[CrossRef]
    [Google Scholar]
  27. Suerbaum, S., Josenhans, C., Sterzenbach, T. & 19 other authors ( 2003; ). The complete genome sequence of the carcinogenic bacterium Helicobacter hepaticus. Proc Natl Acad Sci U S A 100, 7901–7906.[CrossRef]
    [Google Scholar]
  28. Tomb, J. F., White, O., Kerlavage, A. R. & 39 other authors ( 1997; ). The complete genome sequence of the gastric pathogen Helicobacter pylori. Nature 388, 539–547.[CrossRef]
    [Google Scholar]
  29. Tsuda, M., Karita, M., Morshed, M. G., Okita, K. & Nakazawa, T. ( 1994; ). A urease-negative mutant of Helicobacter pylori constructed by allelic exchange mutagenesis lacks the ability to colonize the nude mouse stomach. Infect Immun 62, 3586–3589.
    [Google Scholar]
  30. van Vliet, A. H. M., Wooldridge, K. G. & Ketley, J. M. ( 1998; ). Iron-responsive gene regulation in a Campylobacter jejuni fur mutant. J Bacteriol 180, 5291–5298.
    [Google Scholar]
  31. van Vliet, A. H. M., Kuipers, E. J., Waidner, B. & 7 other authors ( 2001; ). Nickel-responsive induction of urease expression in Helicobacter pylori is mediated at the transcriptional level. Infect Immun 69, 4891–4897.[CrossRef]
    [Google Scholar]
  32. van Vliet, A. H. M., Poppelaars, S. W., Davies, B. J., Stoof, J., Bereswill, S., Kist, M., Penn, C. W., Kuipers, E. J. & Kusters, J. G. ( 2002; ). NikR mediates nickel-responsive transcriptional induction of urease expression in Helicobacter pylori. Infect Immun 70, 2846–2852.[CrossRef]
    [Google Scholar]
  33. van Vliet, A. H. M., Ernst, F. D. & Kusters, J. G. ( 2004a; ). NikR-mediated regulation of Helicobacter pylori acid adaptation. Trends Microbiol 12, 489–494.[CrossRef]
    [Google Scholar]
  34. van Vliet, A. H. M., Kuipers, E. J., Stoof, J., Poppelaars, S. W. & Kusters, J. G. ( 2004b; ). Acid-responsive gene induction of ammonia-producing enzymes in Helicobacter pylori is mediated via a metal-responsive repressor cascade. Infect Immun 72, 766–773.[CrossRef]
    [Google Scholar]
  35. Verhoef, C., Pot, R. G. J., de Man, R. A., Zondervan, P. E., Kuipers, E. J., IJzermans, J. N. & Kusters, J. G. ( 2003; ). Detection of identical Helicobacter DNA in the stomach and in the non-cirrhotic liver of patients with hepatocellular carcinoma. Eur J Gastroenterol Hepatol 15, 1171–1174.[CrossRef]
    [Google Scholar]
  36. Ward, J. M., Fox, J. G., Anver, M. R. & 7 other authors ( 1994; ). Chronic active hepatitis and associated liver tumors in mice caused by a persistent bacterial infection with a novel Helicobacter species. J Natl Cancer Inst 86, 1222–1227.[CrossRef]
    [Google Scholar]
  37. Weeks, D. L., Gushansky, G., Scott, D. R. & Sachs, G. ( 2004; ). Mechanism of proton gating of a urea channel. J Biol Chem 279, 9944–9950.[CrossRef]
    [Google Scholar]
  38. Williams, C. L., Preston, T., Hossack, M., Slater, C. & McColl, K. E. ( 1996; ). Helicobacter pylori utilises urea for amino acid synthesis. FEMS Immunol Med Microbiol 13, 87–94.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.28188-0
Loading
/content/journal/micro/10.1099/mic.0.28188-0
Loading

Data & Media loading...

does not survive acid shock. ATCC 51449 was incubated for 30 min in PBS of pH 7.0, pH 5.5 and pH 3.0 in either unsupplemented PBS (grey bars), PBS with 0.5 mM urea (white bars), or PBS with 100 µM Ni Cl (black bars). Asterisks at pH 3.0 indicate that there was no survival of at this condition. Data points shown are the mean of three independent experiments. [PDF](11 kb) Urease activity is nickel-induced in . Urease activity of strain ATCC 51449 (a) and strain 26695 (b) was assessed after 24 h of growth at pH 7.0 in unsupplemented BBC medium (black bars) and BBC medium supplemented with NiCl to a final concentration of 100 µM (white bars). Note the difference in the scale of the -axis. Each bar represents data from three to five independent experiments for each strain; error bars denote standard deviation. Asterisks indicate a significant increase of urease activity in the test condition (100 µM Ni Cl ) when compared to the standard growth conditions (unsupplemented medium) ( ≤0.05, Mann-Whitney U test). [PDF](12 kb)

PDF

does not survive acid shock. ATCC 51449 was incubated for 30 min in PBS of pH 7.0, pH 5.5 and pH 3.0 in either unsupplemented PBS (grey bars), PBS with 0.5 mM urea (white bars), or PBS with 100 µM Ni Cl (black bars). Asterisks at pH 3.0 indicate that there was no survival of at this condition. Data points shown are the mean of three independent experiments. [PDF](11 kb) Urease activity is nickel-induced in . Urease activity of strain ATCC 51449 (a) and strain 26695 (b) was assessed after 24 h of growth at pH 7.0 in unsupplemented BBC medium (black bars) and BBC medium supplemented with NiCl to a final concentration of 100 µM (white bars). Note the difference in the scale of the -axis. Each bar represents data from three to five independent experiments for each strain; error bars denote standard deviation. Asterisks indicate a significant increase of urease activity in the test condition (100 µM Ni Cl ) when compared to the standard growth conditions (unsupplemented medium) ( ≤0.05, Mann-Whitney U test). [PDF](12 kb)

PDF
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