1887

Abstract

The gene encoding the manganese-containing superoxide dismutase (MnSOD) of was characterized. It is transcribed monocistronically from an upstream promoter identified by rapid amplification of cDNA ends (RACE)-PCR. A mutant was constructed and characterized. Growth of the mutant strain was not significantly different from that of its wild-type counterpart in standing and aerated cultures. However, the mutant was more sensitive towards menadione and hydroperoxide stresses. The response to HO stress was analysed in more detail, and the mode of killing of this oxidant was different under anaerobic and aerobic conditions. Cultures grown and challenged under anaerobic conditions were highly sensitive to treatment with 35 mM HO. They were largely protected by the iron chelator deferoxamine, which suggested that killing was mainly due to an enhanced Fenton reaction. In contrast, neither strain was protected by the iron chelators deferoxamine and diethylenetriaminepentaacteic acid when grown and challenged under aerobic conditions, which suggested that inactivation of the cells by HO was due to another killing mode. The mutant was more sensitive under these conditions, showing that MnSOD is also important for protecting the cells from damage under aerobic conditions. Finally, the MnSOD of may be considered to be a virulence factor, since survival of the corresponding mutant strain was highly affected inside mouse peritoneal macrophages.

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2006-09-01
2019-12-08
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References

  1. Almiron, M. A. J., Link, D., Furlong, D. & Kolter, R. ( 1992; ). A novel DNA-binding protein with regulatory and protective roles in starved Escherichia coli. Genes Dev 6, 2646–2654.[CrossRef]
    [Google Scholar]
  2. Arnaud, M., Chastanet, A. & Débarbouillé, M. ( 2004; ). A new vector for efficient allelic replacement in naturally non transformable low GC% Gram-positive bacteria. Appl Environ Microbiol 70, 6887–6891.[CrossRef]
    [Google Scholar]
  3. Beauchamp, C. & Fridovich, I. ( 1971; ). Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44, 276–287.[CrossRef]
    [Google Scholar]
  4. Britton, L., Malinowski, D. P. & Fridovich, I. ( 1978; ). Superoxide dismutase and oxygen metabolism in Streptococcus faecalis and comparisons with other organisms. J Bacteriol 134, 229–236.
    [Google Scholar]
  5. Burr, T., Mitchell, J., Kolb, A., Minchin, S. & Busby, S. ( 2000; ). DNA sequence elements located immediately upstream of the −10 hexamer in Escherichia coli promoters: a systematic study. Nucleic Acids Res 28, 1864–1870.[CrossRef]
    [Google Scholar]
  6. Carlioz, A. & Touati, D. ( 1986; ). Isolation of superoxide dismutase mutants in Escherichia coli: is superoxide dismutase necessary for aerobic life? EMBO J 5, 623–630.
    [Google Scholar]
  7. Chang, S. K. & Hassan, H. M. ( 1997; ). Characterisation of superoxide dismutase in Streptococcus thermophilus. Appl Environ Microbiol 63, 3732–3735.
    [Google Scholar]
  8. Farr, S. B., D'Ari, R. & Touati, D. ( 1986; ). Oxygen-dependent mutagenesis in Escherichia coli lacking superoxide dismutase. Proc Natl Acad Sci U S A 83, 8268–8272.[CrossRef]
    [Google Scholar]
  9. Flahaut, S., Laplace, J. M., Frère, J. & Auffray, Y. ( 1998; ). The oxidative stress response in Enterococcus faecalis: relationship between H2O2 tolerance and H2O2 stress proteins. Lett Appl Microbiol 26, 259–264.[CrossRef]
    [Google Scholar]
  10. Gardner, P. R. & Fridovich, I. ( 1992; ). Inactivation-reactivation of aconitase in Escherichia coli. A sensitive measure of superoxide radical. J Biol Chem 267, 8757–8763.
    [Google Scholar]
  11. Gentry-Weeks, C. R., Karkhoff-Schweizer, R., Pikis, A., Estay, M. & Keith, J. M. ( 1999; ). Survival of Enterococcus faecalis in mouse peritoneal macrophages. Infect Immun 67, 2160–2165.
    [Google Scholar]
  12. Giles, S. S., Batinic-Haberle, I., Perfect, J. R. & Cox, G. M. ( 2005; ). Cryptococcus neoformans mitochondrial superoxide dismutase: an essential link between antioxidant function and high-temperature growth. Eukaryot Cell 4, 46–54.[CrossRef]
    [Google Scholar]
  13. Gilmore, M. S., Coburn, P. S., Nallapareddy, S. R. & Murray, B. E. ( 2002; ). Enterococcal virulence. In The Enterococci: Pathogenesis, Molecular Biology, and Antibiotic Resistance, pp. 301–354. Edited by M. S. Gilmore. Washington, DC: American Society for Microbiology.
  14. Grant, R. A., Filman, D. J., Finkel, S. E., Kolter, R. & Hogle, J. M. ( 1998; ). The crystal structure of Dps, a ferritin homolog that binds and protects DNA. Nat Struct Biol 5, 294–303.[CrossRef]
    [Google Scholar]
  15. Gregory, E. M. & Fridovich, I. ( 1973; ). Induction of superoxide dismutase by molecular oxygen. J Bacteriol 114, 543–548.
    [Google Scholar]
  16. Hassan, H. M. ( 1989; ). Microbial superoxide dismutases. Adv Genet 26, 65–97.
    [Google Scholar]
  17. Hassett, D. J. & Cohen, M. S. ( 1989; ). Bacterial adaptation to oxidative stress: implications for pathogenesis and interaction with phagocytic cells. FASEB J 3, 2574–2582.
    [Google Scholar]
  18. Huycke, M. M., Joyce, W. & Wack, M. F. ( 1996; ). Augmented production of extracellular superoxide by blood isolates of Enterococcus faecalis. J Infect Dis 173, 743–746.[CrossRef]
    [Google Scholar]
  19. Huycke, M. M., Moore, D., Joyce, W., Wise, P., Shepard, L., Kotake, Y. & Gilmore, M. S. ( 2001; ). Extracellular superoxide production by Enterococcus faecalis requires demethylmenaquinone and is attenuated by functional terminal quinol oxidases. Mol Microbiol 42, 729–740.
    [Google Scholar]
  20. Imlay, J. A. ( 2002; ). How oxygen damages microbes: oxygen tolerance and obligate anaerobiosis. Adv Microb Physiol 46, 111–153.
    [Google Scholar]
  21. Imlay, J. A. ( 2003; ). Pathways of oxidative damage. Annu Rev Microbiol 57, 395–418.[CrossRef]
    [Google Scholar]
  22. Imlay, J. A. & Linn, S. ( 1986; ). Bimodal pattern of killing of DNA-repair-defective or anaerobically grown Escherichia coli by hydrogen peroxide. J Bacteriol 166, 519–527.
    [Google Scholar]
  23. Imlay, J. A. & Linn, S. ( 1988; ). Toxic DNA damage by hydrogen peroxide through the Fenton reaction in vivo and in vitro. Science 240, 640–642.[CrossRef]
    [Google Scholar]
  24. Inaoka, T., Matsumura, Y. & Tsuchido, T. ( 1998; ). Molecular cloning and nucleotide sequence of the superoxide dismutase gene and characterization of its product from Bacillus subtilis. J Bacteriol 180, 3697–3703.
    [Google Scholar]
  25. Jacob, A. E. & Hobbs, S. J. ( 1974; ). Conjugal transfer of plasmid-borne multiple antibiotic resistance in Streptococcus faecalis var. zymogenes. J Bacteriol 117, 360–372.
    [Google Scholar]
  26. Jett, B. D., Huycke, M. M. & Gilmore, M. S. ( 1994; ). Virulence of enterococci. Clin Microbiol Rev 7, 462–478.
    [Google Scholar]
  27. Kak, V. & Chow, J. W. ( 2002; ). Acquired antibiotic resistances in Enterococci. In The Enterococci: Pathogenesis, Molecular Biology, and Antibiotic Resistance, pp. 355–383. Edited by M. S. Gilmore. Washington, DC: American Society for Microbiology.
  28. Keyer, K. & Imlay, J. A. ( 1996; ). Superoxide accelerates DNA damage by elevating free-iron levels. Proc Natl Acad Sci U S A 93, 13635–13640.[CrossRef]
    [Google Scholar]
  29. Keyer, K., Gort, A. S. & Imlay, J. A. ( 1995; ). Superoxide and the production of oxidative DNA damage. J Bacteriol 177, 6782–6790.
    [Google Scholar]
  30. Laemmli, U. K. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.[CrossRef]
    [Google Scholar]
  31. Law, J., Buist, G., Haandrikman, A., Kok, J., Venema, G. & Leenhouts, K. ( 1995; ). A system to generate chromosomal mutations in Lactococcus lactis allows fast analysis of targeted genes. J Bacteriol 177, 7011–7018.
    [Google Scholar]
  32. Lowry, O. H., Rosebrough, N. J., Farr, A. L. & Randall, R. J. ( 1951; ). Protein measurement with Folin phenol reagent. J Biol Chem 193, 265–275.
    [Google Scholar]
  33. Maguin, E., Duwat, P., Hege, T. & Ehrlich, D. ( 1992; ). New thermosensitive plasmid for Gram-positive bacteria. J Bacteriol 174, 5633–5638.
    [Google Scholar]
  34. Narasipura, S. D., Ault, J. G., Behr, M. J., Chaturvedi, V. & Chaturvedi, S. ( 2003; ). Characterization of Cu, Zn superoxide dismutase (SOD1) gene knock-out mutant of Cryptococcus neoformans var. gattii: role in biology and virulence. Mol Microbiol 47, 1681–1694.[CrossRef]
    [Google Scholar]
  35. Narasipura, S. D., Chaturvedi, V. & Chaturvedi, S. ( 2005; ). Characterization of Cryptococcus neoformans variety gattii SOD2 reveals distinct roles of the two superoxide dismutases in fungal biology and virulence. Mol Microbiol 55, 1782–1800.[CrossRef]
    [Google Scholar]
  36. Parker M. W. & Blake C. C. ( 1988; ). Iron and manganese containing superoxide-dismutases can be distinguished by analysis of their primary structures. FEBS Lett 229, 377–382.
  37. Pedras Vasconcelos, J. A. & Deneer, H. G. ( 1994; ). Expression of superoxide dismutase in Listeria monocytogenes. Appl Environ Microbiol 60, 2360–2366.
    [Google Scholar]
  38. Poyart, C., Pellegrini, E., Guillot, O., Boumaila, C., Baptista, M. & Trieu-Cuot, P. ( 2001; ). Contribution of Mn-cofactored superoxide dismutase (SODA) to the virulence of Streptococcus agalactiae. Infect Immun 69, 5098–5106.[CrossRef]
    [Google Scholar]
  39. Sambrook, J., Fritsch, E. & Maniatis, T. ( 1989; ). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  40. Terzaghi, B. E. & Sandine, W. ( 1975; ). Improved medium for lactic streptococci and their bacteriophages. Appl Environ Microbiol 29, 807–813.
    [Google Scholar]
  41. Touati, D. ( 2002; ). Investigating phenotypes resulting from a lack of superoxide dismutase in bacterial null mutants. Methods Enzymol 349, 145–154.
    [Google Scholar]
  42. Verneuil, N., Sanguinetti, M., Le Breton, Y., Posteraro, B., Fadda, G., Auffray, Y., Hartke, A. & Giard, J.-C. ( 2004; ). Effects of Enterococcus faecalis hypR gene encoding a new transcriptional regulator on oxidative stress response and intracellular survival within macrophages. Infect Immun 72, 4424–4431.[CrossRef]
    [Google Scholar]
  43. Verneuil, N., Rincé, A., Sanguinetti, M., Auffray, Y., Hartke, A. & Giard, J.-C. ( 2005; ). Implication of hypR in virulence and oxidative stress response of Enterococcus faecalis. FEMS Microbiol Lett 252, 137–141.[CrossRef]
    [Google Scholar]
  44. Welch, K. D., Davis, T. Z. & Aust, S. D. ( 2002; ). Iron autooxidation and free radical generation: effects of buffers, ligands, and chelators. Arch Biochem Biophys 397, 360–369.[CrossRef]
    [Google Scholar]
  45. Yagi, Y. & Clewell, D. B. ( 1980; ). Recombination-deficient mutant of Streptococcus faecalis. J Bacteriol 143, 966–970.
    [Google Scholar]
  46. Yesilkaya, H., Kadioglu, A., Gingles, N., Alexander, J. E., Mitchell, T. & Andrew, P. W. ( 2000; ). Role of manganese-containing superoxide dismutase in oxidative stress and virulence of Streptococcus pneumoniae. Infect Immun 68, 2819–2856.[CrossRef]
    [Google Scholar]
  47. Zhao, G., Ceci, P., Ilari, A., Giangiacomo, L., Laue, T. M., Chiancone, E. & Chasteen, N. D. ( 2002; ). Iron and hydrogen peroxide detoxification properties of DNA-binding protein from starved cells. A ferritin-like DNA-binding protein of Escherichia coli. J Biol Chem 277, 27689–27696.[CrossRef]
    [Google Scholar]
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