Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Oct;177(20):5924–5929. doi: 10.1128/jb.177.20.5924-5929.1995

Function and stationary-phase induction of periplasmic copper-zinc superoxide dismutase and catalase/peroxidase in Caulobacter crescentus.

S Schnell 1, H M Steinman 1
PMCID: PMC177420  PMID: 7592345

Abstract

Although cytosolic superoxide dismutases (SODs) are widely distributed among bacteria, only a small number of species contain a periplasmic SOD. One of these is Caulobacter crescentus, which has a copper-zinc SOD (CuZnSOD) in the periplasm and an iron SOD (FeSOD) in the cytosol. The function of periplasmic CuZnSOD was studied by characterizing a mutant of C. crescentus with an insertionally inactivated CuZnSOD gene. Wild-type and mutant strains showed identical tolerance to intracellular superoxide. However, in response to extracellular superoxide, the presence of periplasmic CuZnSOD increased survival by as much as 20-fold. This is the first demonstration that periplasmic SOD defends against external superoxide of environmental origin. This result has implications for those bacterial pathogens that contain a CuZnSOD. C. crescentus was shown to contain a single catalase/peroxidase which, like Escherichia coli KatG catalase/peroxidase, is present in both the periplasmic and cytoplasmic fractions. The growth stage dependence of C. crescentus catalase/peroxidase and SOD activity was studied. Although FeSOD activity was identical in exponential- and stationary-phase cultures, CuZnSOD was induced nearly 4-fold in stationary phase and the catalase/peroxidase was induced nearly 100-fold. Induction of antioxidant enzymes in the periplasm of C. crescentus appears to be an important attribute of the stationary-phase response and may be a useful tool for studying its regulation.

Full Text

The Full Text of this article is available as a PDF (232.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Beyer W., Imlay J., Fridovich I. Superoxide dismutases. Prog Nucleic Acid Res Mol Biol. 1991;40:221–253. doi: 10.1016/s0079-6603(08)60843-0. [DOI] [PubMed] [Google Scholar]
  2. Bricker B. J., Tabatabai L. B., Judge B. A., Deyoe B. L., Mayfield J. E. Cloning, expression, and occurrence of the Brucella Cu-Zn superoxide dismutase. Infect Immun. 1990 Sep;58(9):2935–2939. doi: 10.1128/iai.58.9.2935-2939.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Claiborne A., Fridovich I. Purification of the o-dianisidine peroxidase from Escherichia coli B. Physicochemical characterization and analysis of its dual catalatic and peroxidatic activities. J Biol Chem. 1979 May 25;254(10):4245–4252. [PubMed] [Google Scholar]
  5. Clare D. A., Duong M. N., Darr D., Archibald F., Fridovich I. Effects of molecular oxygen on detection of superoxide radical with nitroblue tetrazolium and on activity stains for catalase. Anal Biochem. 1984 Aug 1;140(2):532–537. doi: 10.1016/0003-2697(84)90204-5. [DOI] [PubMed] [Google Scholar]
  6. Colman R. F. Effect of modification of a methionyl residue on the kinetic and molecular properties of isocitrate dehydrogenase. J Biol Chem. 1968 May 25;243(10):2454–2464. [PubMed] [Google Scholar]
  7. Farr S. B., D'Ari R., Touati D. Oxygen-dependent mutagenesis in Escherichia coli lacking superoxide dismutase. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8268–8272. doi: 10.1073/pnas.83.21.8268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Farr S. B., Touati D., Kogoma T. Effects of oxygen stress on membrane functions in Escherichia coli: role of HPI catalase. J Bacteriol. 1988 Apr;170(4):1837–1842. doi: 10.1128/jb.170.4.1837-1842.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fridovich I. Superoxide radical: an endogenous toxicant. Annu Rev Pharmacol Toxicol. 1983;23:239–257. doi: 10.1146/annurev.pa.23.040183.001323. [DOI] [PubMed] [Google Scholar]
  10. Grace S. C. Phylogenetic distribution of superoxide dismutase supports an endosymbiotic origin for chloroplasts and mitochondria. Life Sci. 1990;47(21):1875–1886. doi: 10.1016/0024-3205(90)90399-c. [DOI] [PubMed] [Google Scholar]
  11. Green M. J., Hill H. A. Chemistry of dioxygen. Methods Enzymol. 1984;105:3–22. doi: 10.1016/s0076-6879(84)05004-7. [DOI] [PubMed] [Google Scholar]
  12. Gregory E. M., Fridovich I. Visualization of catalase on acrylamide gels. Anal Biochem. 1974 Mar;58(1):57–62. doi: 10.1016/0003-2697(74)90440-0. [DOI] [PubMed] [Google Scholar]
  13. Hassan H. M. Determination of microbial damage caused by oxygen free radicals, and the protective role of superoxide dismutase. Methods Enzymol. 1984;105:404–412. doi: 10.1016/s0076-6879(84)05056-4. [DOI] [PubMed] [Google Scholar]
  14. Hassan H. M., Fridovich I. Paraquat and Escherichia coli. Mechanism of production of extracellular superoxide radical. J Biol Chem. 1979 Nov 10;254(21):10846–10852. [PubMed] [Google Scholar]
  15. Hassan H. M. Microbial superoxide dismutases. Adv Genet. 1989;26:65–97. doi: 10.1016/s0065-2660(08)60223-0. [DOI] [PubMed] [Google Scholar]
  16. Heimberger A., Eisenstark A. Compartmentalization of catalases in Escherichia coli. Biochem Biophys Res Commun. 1988 Jul 15;154(1):392–397. doi: 10.1016/0006-291x(88)90698-5. [DOI] [PubMed] [Google Scholar]
  17. Hengge-Aronis R. Survival of hunger and stress: the role of rpoS in early stationary phase gene regulation in E. coli. Cell. 1993 Jan 29;72(2):165–168. doi: 10.1016/0092-8674(93)90655-a. [DOI] [PubMed] [Google Scholar]
  18. Hopkin K. A., Papazian M. A., Steinman H. M. Functional differences between manganese and iron superoxide dismutases in Escherichia coli K-12. J Biol Chem. 1992 Dec 5;267(34):24253–24258. [PubMed] [Google Scholar]
  19. Ivanova A., Miller C., Glinsky G., Eisenstark A. Role of rpoS (katF) in oxyR-independent regulation of hydroperoxidase I in Escherichia coli. Mol Microbiol. 1994 May;12(4):571–578. doi: 10.1111/j.1365-2958.1994.tb01043.x. [DOI] [PubMed] [Google Scholar]
  20. Johnson R. C., Ely B. Analysis of nonmotile mutants of the dimorphic bacterium Caulobacter crescentus. J Bacteriol. 1979 Jan;137(1):627–634. doi: 10.1128/jb.137.1.627-634.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kargalioglu Y., Imlay J. A. Importance of anaerobic superoxide dismutase synthesis in facilitating outgrowth of Escherichia coli upon entry into an aerobic habitat. J Bacteriol. 1994 Dec;176(24):7653–7658. doi: 10.1128/jb.176.24.7653-7658.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kim Y., Watrud L. S., Matin A. A carbon starvation survival gene of Pseudomonas putida is regulated by sigma 54. J Bacteriol. 1995 Apr;177(7):1850–1859. doi: 10.1128/jb.177.7.1850-1859.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kolter R., Siegele D. A., Tormo A. The stationary phase of the bacterial life cycle. Annu Rev Microbiol. 1993;47:855–874. doi: 10.1146/annurev.mi.47.100193.004231. [DOI] [PubMed] [Google Scholar]
  24. Kroll J. S., Langford P. R., Loynds B. M. Copper-zinc superoxide dismutase of Haemophilus influenzae and H. parainfluenzae. J Bacteriol. 1991 Dec;173(23):7449–7457. doi: 10.1128/jb.173.23.7449-7457.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Latimer E., Simmers J., Sriranganathan N., Roop R. M., 2nd, Schurig G. G., Boyle S. M. Brucella abortus deficient in copper/zinc superoxide dismutase is virulent in BALB/c mice. Microb Pathog. 1992 Feb;12(2):105–113. doi: 10.1016/0882-4010(92)90113-3. [DOI] [PubMed] [Google Scholar]
  26. Lavelle F., Michelson A. M., Dimitrijevic L. Biological protection by superoxide dismutase. Biochem Biophys Res Commun. 1973 Nov 16;55(2):350–357. doi: 10.1016/0006-291x(73)91094-2. [DOI] [PubMed] [Google Scholar]
  27. Loewen P. C., Switala J., Triggs-Raine B. L. Catalases HPI and HPII in Escherichia coli are induced independently. Arch Biochem Biophys. 1985 Nov 15;243(1):144–149. doi: 10.1016/0003-9861(85)90782-9. [DOI] [PubMed] [Google Scholar]
  28. Marklund S., Marklund G. Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem. 1974 Sep 16;47(3):469–474. doi: 10.1111/j.1432-1033.1974.tb03714.x. [DOI] [PubMed] [Google Scholar]
  29. Nakayama K. Rapid viability loss on exposure to air in a superoxide dismutase-deficient mutant of Porphyromonas gingivalis. J Bacteriol. 1994 Apr;176(7):1939–1943. doi: 10.1128/jb.176.7.1939-1943.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. POINDEXTER J. S. BIOLOGICAL PROPERTIES AND CLASSIFICATION OF THE CAULOBACTER GROUP. Bacteriol Rev. 1964 Sep;28:231–295. doi: 10.1128/br.28.3.231-295.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Prohaska J. R. Changes in tissue growth, concentrations of copper, iron, cytochrome oxidase and superoxide dismutase subsequent to dietary or genetic copper deficiency in mice. J Nutr. 1983 Oct;113(10):2048–2058. doi: 10.1093/jn/113.10.2048. [DOI] [PubMed] [Google Scholar]
  32. Puget K., Michelson A. M. Isolation of a new copper-containing superoxide dismutase bacteriocuprein. Biochem Biophys Res Commun. 1974 Jun 4;58(3):830–838. doi: 10.1016/s0006-291x(74)80492-4. [DOI] [PubMed] [Google Scholar]
  33. Sadosky A. B., Wilson J. W., Steinman H. M., Shuman H. A. The iron superoxide dismutase of Legionella pneumophila is essential for viability. J Bacteriol. 1994 Jun;176(12):3790–3799. doi: 10.1128/jb.176.12.3790-3799.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  35. Sriranganathan N., Boyle S. M., Schurig G., Misra H. Superoxide dismutases of virulent and avirulent strains of Brucella abortus. Vet Microbiol. 1991 Feb 15;26(4):359–366. doi: 10.1016/0378-1135(91)90029-f. [DOI] [PubMed] [Google Scholar]
  36. Stabel T. J., Sha Z., Mayfield J. E. Periplasmic location of Brucella abortus Cu/Zn superoxide dismutase. Vet Microbiol. 1994 Feb;38(4):307–314. doi: 10.1016/0378-1135(94)90149-x. [DOI] [PubMed] [Google Scholar]
  37. Steinman H. M. Bacteriocuprein superoxide dismutase of Photobacterium leiognathi. Isolation and sequence of the gene and evidence for a precursor form. J Biol Chem. 1987 Feb 5;262(4):1882–1887. [PubMed] [Google Scholar]
  38. Steinman H. M. Bacteriocuprein superoxide dismutases in pseudomonads. J Bacteriol. 1985 Jun;162(3):1255–1260. doi: 10.1128/jb.162.3.1255-1260.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Steinman H. M. Construction of an Escherichia coli K-12 strain deleted for manganese and iron superoxide dismutase genes and its use in cloning the iron superoxide dismutase gene of Legionella pneumophila. Mol Gen Genet. 1992 Apr;232(3):427–430. doi: 10.1007/BF00266247. [DOI] [PubMed] [Google Scholar]
  40. Steinman H. M., Ely B. Copper-zinc superoxide dismutase of Caulobacter crescentus: cloning, sequencing, and mapping of the gene and periplasmic location of the enzyme. J Bacteriol. 1990 Jun;172(6):2901–2910. doi: 10.1128/jb.172.6.2901-2910.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Steinman H. M. Function of periplasmic copper-zinc superoxide dismutase in Caulobacter crescentus. J Bacteriol. 1993 Feb;175(4):1198–1202. doi: 10.1128/jb.175.4.1198-1202.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Steinman H. M., Weinstein L., Brenowitz M. The manganese superoxide dismutase of Escherichia coli K-12 associates with DNA. J Biol Chem. 1994 Nov 18;269(46):28629–28634. [PubMed] [Google Scholar]
  43. Tatum F. M., Detilleux P. G., Sacks J. M., Halling S. M. Construction of Cu-Zn superoxide dismutase deletion mutants of Brucella abortus: analysis of survival in vitro in epithelial and phagocytic cells and in vivo in mice. Infect Immun. 1992 Jul;60(7):2863–2869. doi: 10.1128/iai.60.7.2863-2869.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Witholt B., Boekhout M., Brock M., Kingma J., Heerikhuizen H. V., Leij L. D. An efficient and reproducible procedure for the formation of spheroplasts from variously grown Escherichia coli. Anal Biochem. 1976 Jul;74(1):160–170. doi: 10.1016/0003-2697(76)90320-1. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES