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. 1995 May;63(5):1739–1744. doi: 10.1128/iai.63.5.1739-1744.1995

Role of Salmonella typhimurium Mn-superoxide dismutase (SodA) in protection against early killing by J774 macrophages.

R M Tsolis 1, A J Bäumler 1, F Heffron 1
PMCID: PMC173218  PMID: 7729880

Abstract

The Salmonella typhimurium gene for Mn-cofactored superoxide dismutase (sodA) was cloned by complementation of an Escherichia coli sodA sodB mutant for growth on minimal medium. Sequence analysis revealed an open reading frame of 618 bp encoding a polypeptide with 97% identity to E. coli SodA. A S. typhimurium sodA mutant was created by allelic exchange and tested for the ability to survive in the murine macrophage-like cell line J774. Growth of bacteria under iron-limiting conditions, inactivation of the Fur repressor, or expression of sodA from a plasmid resulted in increased resistance to early killing by J774 cells, which was abolished in the sodA mutant. These results suggest that resistance to the early oxygen-dependent microbicidal mechanisms of phagocytes involves the SodA gene product. The S. typhimurium sodA mutant was not significantly attenuated in mice, however, which suggests that resistance to early oxygen-dependent microbicidal mechanisms in vivo may play only a minor role in Salmonella pathogenesis.

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Selected References

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  1. Beaman L., Beaman B. L. The role of oxygen and its derivatives in microbial pathogenesis and host defense. Annu Rev Microbiol. 1984;38:27–48. doi: 10.1146/annurev.mi.38.100184.000331. [DOI] [PubMed] [Google Scholar]
  2. Beauchamp C., Fridovich I. Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem. 1971 Nov;44(1):276–287. doi: 10.1016/0003-2697(71)90370-8. [DOI] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Carlioz A., Touati D. Isolation of superoxide dismutase mutants in Escherichia coli: is superoxide dismutase necessary for aerobic life? EMBO J. 1986 Mar;5(3):623–630. doi: 10.1002/j.1460-2075.1986.tb04256.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Compan I., Touati D. Interaction of six global transcription regulators in expression of manganese superoxide dismutase in Escherichia coli K-12. J Bacteriol. 1993 Mar;175(6):1687–1696. doi: 10.1128/jb.175.6.1687-1696.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Damiani G., Kiyotaki C., Soeller W., Sasada M., Peisach J., Bloom B. R. Macrophage variants in oxygen metabolism. J Exp Med. 1980 Oct 1;152(4):808–822. doi: 10.1084/jem.152.4.808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Farr S. B., Kogoma T. Oxidative stress responses in Escherichia coli and Salmonella typhimurium. Microbiol Rev. 1991 Dec;55(4):561–585. doi: 10.1128/mr.55.4.561-585.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Franzon V. L., Arondel J., Sansonetti P. J. Contribution of superoxide dismutase and catalase activities to Shigella flexneri pathogenesis. Infect Immun. 1990 Feb;58(2):529–535. doi: 10.1128/iai.58.2.529-535.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Garcia-del Portillo F., Foster J. W., Maguire M. E., Finlay B. B. Characterization of the micro-environment of Salmonella typhimurium-containing vacuoles within MDCK epithelial cells. Mol Microbiol. 1992 Nov;6(22):3289–3297. doi: 10.1111/j.1365-2958.1992.tb02197.x. [DOI] [PubMed] [Google Scholar]
  10. Hassett D. J., Cohen M. S. Bacterial adaptation to oxidative stress: implications for pathogenesis and interaction with phagocytic cells. FASEB J. 1989 Dec;3(14):2574–2582. doi: 10.1096/fasebj.3.14.2556311. [DOI] [PubMed] [Google Scholar]
  11. Ishibashi Y., Arai T. Roles of the complement receptor type 1 (CR1) and type 3 (CR3) on phagocytosis and subsequent phagosome-lysosome fusion in Salmonella-infected murine macrophages. FEMS Microbiol Immunol. 1990 Sep;2(2):89–96. doi: 10.1111/j.1574-6968.1990.tb03505.x. [DOI] [PubMed] [Google Scholar]
  12. Kinder S. A., Badger J. L., Bryant G. O., Pepe J. C., Miller V. L. Cloning of the YenI restriction endonuclease and methyltransferase from Yersinia enterocolitica serotype O8 and construction of a transformable R-M+ mutant. Gene. 1993 Dec 22;136(1-2):271–275. doi: 10.1016/0378-1119(93)90478-l. [DOI] [PubMed] [Google Scholar]
  13. Kossack R. E., Guerrant R. L., Densen P., Schadelin J., Mandell G. L. Diminished neutrophil oxidative metabolism after phagocytosis of virulent Salmonella typhi. Infect Immun. 1981 Feb;31(2):674–678. doi: 10.1128/iai.31.2.674-678.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kraft R., Tardiff J., Krauter K. S., Leinwand L. A. Using mini-prep plasmid DNA for sequencing double stranded templates with Sequenase. Biotechniques. 1988 Jun;6(6):544-6, 549. [PubMed] [Google Scholar]
  15. Law S. K. C3 receptors on macrophages. J Cell Sci Suppl. 1988;9:67–97. doi: 10.1242/jcs.1988.supplement_9.4. [DOI] [PubMed] [Google Scholar]
  16. Lepay D. A., Nathan C. F., Steinman R. M., Murray H. W., Cohn Z. A. Murine Kupffer cells. Mononuclear phagocytes deficient in the generation of reactive oxygen intermediates. J Exp Med. 1985 May 1;161(5):1079–1096. doi: 10.1084/jem.161.5.1079. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Libby S. J., Goebel W., Ludwig A., Buchmeier N., Bowe F., Fang F. C., Guiney D. G., Songer J. G., Heffron F. A cytolysin encoded by Salmonella is required for survival within macrophages. Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):489–493. doi: 10.1073/pnas.91.2.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miller R. M., Garbus J., Hornick R. B. Lack of enhanced oxygen consumption by polymorphonuclear leukocytes on phagocytosis of virulent Salmonella typhi. Science. 1972 Mar 3;175(4025):1010–1011. doi: 10.1126/science.175.4025.1010. [DOI] [PubMed] [Google Scholar]
  19. Niederhoffer E. C., Naranjo C. M., Bradley K. L., Fee J. A. Control of Escherichia coli superoxide dismutase (sodA and sodB) genes by the ferric uptake regulation (fur) locus. J Bacteriol. 1990 Apr;172(4):1930–1938. doi: 10.1128/jb.172.4.1930-1938.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. North R. J. The relative importance of blood monocytes and fixed macrophages to the expression of cell-mediated immunity to infection. J Exp Med. 1970 Sep 1;132(3):521–534. doi: 10.1084/jem.132.3.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Papp-Szabò E., Firtel M., Josephy P. D. Comparison of the sensitivities of Salmonella typhimurium oxyR and katG mutants to killing by human neutrophils. Infect Immun. 1994 Jul;62(7):2662–2668. doi: 10.1128/iai.62.7.2662-2668.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Papp-Szabò E., Sutherland C. L., Josephy P. D. Superoxide dismutase and the resistance of Escherichia coli to phagocytic killing by human neutrophils. Infect Immun. 1993 Apr;61(4):1442–1446. doi: 10.1128/iai.61.4.1442-1446.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Payne N. R., Horwitz M. A. Phagocytosis of Legionella pneumophila is mediated by human monocyte complement receptors. J Exp Med. 1987 Nov 1;166(5):1377–1389. doi: 10.1084/jem.166.5.1377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schlesinger L. S., Horwitz M. A. Phagocytosis of Mycobacterium leprae by human monocyte-derived macrophages is mediated by complement receptors CR1 (CD35), CR3 (CD11b/CD18), and CR4 (CD11c/CD18) and IFN-gamma activation inhibits complement receptor function and phagocytosis of this bacterium. J Immunol. 1991 Sep 15;147(6):1983–1994. [PubMed] [Google Scholar]
  25. Stojiljkovic I., Bäumler A. J., Heffron F. Ethanolamine utilization in Salmonella typhimurium: nucleotide sequence, protein expression, and mutational analysis of the cchA cchB eutE eutJ eutG eutH gene cluster. J Bacteriol. 1995 Mar;177(5):1357–1366. doi: 10.1128/jb.177.5.1357-1366.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Touati D. Cloning and mapping of the manganese superoxide dismutase gene (sodA) of Escherichia coli K-12. J Bacteriol. 1983 Sep;155(3):1078–1087. doi: 10.1128/jb.155.3.1078-1087.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Van Camp W., Bowler C., Villarroel R., Tsang E. W., Van Montagu M., Inzé D. Characterization of iron superoxide dismutase cDNAs from plants obtained by genetic complementation in Escherichia coli. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9903–9907. doi: 10.1073/pnas.87.24.9903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Vladoianu I. R., Chang H. R., Pechère J. C. Expression of host resistance to Salmonella typhi and Salmonella typhimurium: bacterial survival within macrophages of murine and human origin. Microb Pathog. 1990 Feb;8(2):83–90. doi: 10.1016/0882-4010(90)90072-x. [DOI] [PubMed] [Google Scholar]

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