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Volume 158, Issue 8

Roles of H uptake hydrogenases in acid tolerance Free

Abstract

Hydrogenases play many roles in bacterial physiology, and use of H by the uptake-type enzymes of animal pathogens is of particular interest. Hydrogenases have never been studied in the pathogen , so targeted mutant strains were individually generated in the two H-uptake enzymes (Hya and Hyb) and in the H-evolving enzyme (Hyc) to address their roles. Under anaerobic fermentative conditions, a Hya mutant strain () was unable to oxidize H, while a Hyb mutant strain oxidized H like the wild-type. A strain oxidized more exogenously added hydrogen than the parent. Fluorescence ratio imaging with dye JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide) showed that the parent strain generated a membrane potential 15 times greater than . The mutant was also by far the most acid-sensitive strain, being even more acid-sensitive than a mutant strain in the known acid-combating glutamate-dependent acid-resistance pathway (GDAR pathway). In severe acid-challenge experiments, the addition of glutamate to restored survivability, and this ability was attributed in part to the GDAR system (removes intracellular protons) by mutant strain (e.g. double mutant) analyses. However, mutant strain phenotypes indicated that a larger portion of the glutamate-rescued acid tolerance was independent of GadBC. The acid tolerance of the strains was aided by adding chloride ions to the growth medium. The whole-cell Hya enzyme became more active upon acid exposure (20 min), based on assays of . Indeed, the very high rates of H oxidation by Hya in acid can supply each cell with 2.4×10 protons min. Electrons generated from Hya-mediated H oxidation at the inner membrane likely counteract cytoplasmic positive charge stress, while abundant proton pools deposited periplasmically likely repel proton influx during severe acid stress.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
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2012-08-01
2024-05-09
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References

  1. Brøndsted L., Atlung T. ( 1994). Anaerobic regulation of the hydrogenase 1 (hya) operon of Escherichia coli . J Bacteriol 176:5423–5428[PubMed]
     [Google Scholar]
  2. Castanie-Cornet M. P., Penfound T. A., Smith D., Elliott J. F., Foster J. W. ( 1999). Control of acid resistance in Escherichia coli . J Bacteriol 181:3525–3535[PubMed]
     [Google Scholar]
  3. Cherepanov P. P., Wackernagel W. ( 1995). Gene disruption in Escherichia coli: TcR and KmR cassettes with the option of Flp-catalyzed excision of the antibiotic-resistance determinant. Gene 158:9–14 [View Article][PubMed]
     [Google Scholar]
  4. Coldewey S. M., Hartmann M., Schmidt D. S., Engelking U., Ukena S. N., Gunzer F. ( 2007). Impact of the rpoS genotype for acid resistance patterns of pathogenic and probiotic Escherichia coli . BMC Microbiol 7:21 [View Article][PubMed]
     [Google Scholar]
  5. Datsenko K. A., Wanner B. L. ( 2000). One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 97:6640–6645 [View Article][PubMed]
     [Google Scholar]
  6. Fortier A. H., Leiby D. A., Narayanan R. B., Asafoadjei E., Crawford R. M., Nacy C. A., Meltzer M. S. ( 1995). Growth of Francisella tularensis LVS in macrophages: the acidic intracellular compartment provides essential iron required for growth. Infect Immun 63:1478–1483[PubMed]
     [Google Scholar]
  7. Foster J. W. ( 2004). Escherichia coli acid resistance: tales of an amateur acidophile. Nat Rev Microbiol 2:898–907 [View Article][PubMed]
     [Google Scholar]
  8. Gut H., Pennacchietti E., John R. A., Bossa F., Capitani G., De Biase D., Grütter M. G. ( 2006). Escherichia coli acid resistance: pH-sensing, activation by chloride and autoinhibition in GadB. EMBO J 25:2643–2651 [View Article][PubMed]
     [Google Scholar]
  9. Hung K. F., Byrd J. J., Bose J. L., Kaspar C. W. ( 2006). Reduction of acid tolerance by tetracycline in Escherichia coli expressing tetA(C) is reversed by cations. Appl Environ Microbiol 72:4472–4474 [View Article][PubMed]
     [Google Scholar]
  10. Ingersoll M. A., Zychlinsky A. ( 2006). ShiA abrogates the innate T-cell response to Shigella flexneri infection. Infect Immun 74:2317–2327 [View Article][PubMed]
     [Google Scholar]
  11. Iyer R., Iverson T. M., Accardi A., Miller C. ( 2002). A biological role for prokaryotic ClC chloride channels. Nature 419:715–718 [View Article][PubMed]
     [Google Scholar]
  12. Jennison A. V., Verma N. K. ( 2007). The acid-resistance pathways of Shigella flexneri 2457T. Microbiology 153:2593–2602 [View Article][PubMed]
     [Google Scholar]
  13. Jovanovic G., Lloyd L. J., Stumpf M. P., Mayhew A. J., Buck M. ( 2006). Induction and function of the phage shock protein extracytoplasmic stress response in Escherichia coli . J Biol Chem 281:21147–21161 [View Article][PubMed]
     [Google Scholar]
  14. King P. W., Przybyla A. E. ( 1999). Response of hya expression to external pH in Escherichia coli . J Bacteriol 181:5250–5256[PubMed]
     [Google Scholar]
  15. Lamichhane-Khadka R., Kwiatkowski A., Maier R. J. ( 2010). The Hyb hydrogenase permits hydrogen-dependent respiratory growth of Salmonella enterica serovar Typhimurium. MBio 1:e00284–e10 [View Article][PubMed]
     [Google Scholar]
  16. Lengeler J., Drews G., Schlegel H. (editors) ( 1999). Biology of the Proykaryotes, 1st edn. New York: Wiley-Blackwell;
     [Google Scholar]
  17. Lin J., Lee I. S., Frey J., Slonczewski J. L., Foster J. W. ( 1995). Comparative analysis of extreme acid survival in Salmonella typhimurium, Shigella flexneri, and Escherichia coli . J Bacteriol 177:4097–4104[PubMed]
     [Google Scholar]
  18. Maier R. J. ( 2003). Availability and use of molecular hydrogen as an energy substrate for Helicobacter species. Microbes Infect 5:1159–1163 [View Article][PubMed]
     [Google Scholar]
  19. Maier R. J., Olson J., Olczak A. ( 2003). Hydrogen-oxidizing capabilities of Helicobacter hepaticus and in vivo availability of the substrate. J Bacteriol 185:2680–2682 [View Article][PubMed]
     [Google Scholar]
  20. Maier R. J., Olczak A., Maier S., Soni S., Gunn J. ( 2004). Respiratory hydrogen use by Salmonella enterica serovar Typhimurium is essential for virulence. Infect Immun 72:6294–6299 [View Article][PubMed]
     [Google Scholar]
  21. 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 [View Article][PubMed]
     [Google Scholar]
  22. Noguchi K., Riggins D. P., Eldahan K. C., Kitko R. D., Slonczewski J. L. ( 2010). Hydrogenase-3 contributes to anaerobic acid resistance of Escherichia coli . PLoS ONE 5:e10132 [View Article][PubMed]
     [Google Scholar]
  23. Park K. R., Giard J. C., Eom J. H., Bearson S., Foster J. W. ( 1999). Cyclic AMP receptor protein and TyrR are required for acid pH and anaerobic induction of hyaB and aniC in Salmonella typhimurium . J Bacteriol 181:689–694[PubMed]
     [Google Scholar]
  24. Philpott D. J., Edgeworth J. D., Sansonetti P. J. ( 2000). The pathogenesis of Shigella flexneri infection: lessons from in vitro and in vivo studies. Philos Trans R Soc Lond B Biol Sci 355:575–586 [View Article][PubMed]
     [Google Scholar]
  25. Redwood M. D., Mikheenko I. P., Sargent F., Macaskie L. E. ( 2008). Dissecting the roles of Escherichia coli hydrogenases in biohydrogen production. FEMS Microbiol Lett 278:48–55 [View Article][PubMed]
     [Google Scholar]
  26. Richard H., Foster J. W. ( 2004). Escherichia coli glutamate- and arginine-dependent acid resistance systems increase internal pH and reverse transmembrane potential. J Bacteriol 186:6032–6041 [View Article][PubMed]
     [Google Scholar]
  27. Richard H., Foster J. W. ( 2007). Sodium regulates Escherichia coli acid resistance, and influences GadX- and GadW-dependent activation of gadE . Microbiology 153:3154–3161 [View Article][PubMed]
     [Google Scholar]
  28. Sawers R. G. ( 2005). Formate and its role in hydrogen production in Escherichia coli . Biochem Soc Trans 33:42–46 [View Article][PubMed]
     [Google Scholar]
  29. Trchounian K., Trchounian A. ( 2009). Hydrogenase 2 is most and hydrogenase 1 is less responsible for H2 production by Escherichia coli under glycerol fermentation at neutral and slightly alkaline pH. Int J Hydrogen Energy 34:8839–8845 [View Article]
     [Google Scholar]
  30. Trchounian K., Pinske C., Sawers R. G., Trchounian A. ( 2012). Characterization of Escherichia coli [NiFe]-hydrogenase distribution during fermentative growth at different pHs. Cell Biochem Biophys 62:433–440 [View Article][PubMed]
     [Google Scholar]
  31. Vignais P. M., Colbeau A. ( 2004). Molecular biology of microbial hydrogenases. Curr Issues Mol Biol 6:159–188[PubMed]
     [Google Scholar]
  32. Zbell A. L., Maier R. J. ( 2009). Role of the Hya hydrogenase in recycling of anaerobically produced H2 in Salmonella enterica serovar Typhimurium. Appl Environ Microbiol 75:1456–1459 [View Article][PubMed]
     [Google Scholar]
  33. Zbell A. L., Maier S. E., Maier R. J. ( 2008). Salmonella enterica serovar Typhimurium NiFe uptake-type hydrogenases are differentially expressed in vivo. Infect Immun 76:4445–4454 [View Article][PubMed]
     [Google Scholar]
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Roles of H2 uptake hydrogenases in Shigella flexneri acid tolerance
Microbiology 158, 2204 (2012); https://doi.org/10.1099/mic.0.058248-0
/content/journal/micro/10.1099/mic.0.058248-0
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