Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 Aug;178(15):4688–4695. doi: 10.1128/jb.178.15.4688-4695.1996

Insertional inactivation of Streptococcus pyogenes sod suggests that prtF is regulated in response to a superoxide signal.

C M Gibson 1, M G Caparon 1
PMCID: PMC178240  PMID: 8755901

Abstract

In establishing an infection, Streptococcus pyogenes has the capacity to bind to the host extracellular matrix protein fibronectin via its protein F adhesin. Previous studies have suggested that the expression of protein F is stimulated during aerobic growth or upon addition of superoxide-generating agents to the culture under O2-limited conditions. To further explore the role of superoxide, we have examined the transcription of the gene which encodes protein F (prtF), as well as the expression of superoxide dismutase (SOD) under conditions which promote or repress protein F expression. These studies show that prtF transcription is regulated in response to superoxide concentration and that SOD is regulated in different environments in a manner which directly parallels the expression of protein F. A mutant deficient in SOD activity was constructed by insertional mutation into the gene which encodes SOD (sod). The resulting mutant was sensitive to superoxide and aerobic conditions, showed hypersensitive induction of prtF in response to superoxide, and expressed prtF under normally unfavorable O2-limited conditions. These findings suggest that a streptococcal signal transduction system which senses superoxide may coordinately control expression of prtF and sod.

Full Text

The Full Text of this article is available as a PDF (338.7 KB).

Selected References

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

  1. 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]
  2. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  3. Britton L., Malinowski D. P., Fridovich I. Superoxide dismutase and oxygen metabolism in Streptococcus faecalis and comparisons with other organisms. J Bacteriol. 1978 Apr;134(1):229–236. doi: 10.1128/jb.134.1.229-236.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Caparon M. G., Geist R. T., Perez-Casal J., Scott J. R. Environmental regulation of virulence in group A streptococci: transcription of the gene encoding M protein is stimulated by carbon dioxide. J Bacteriol. 1992 Sep;174(17):5693–5701. doi: 10.1128/jb.174.17.5693-5701.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Caparon M. G., Scott J. R. Genetic manipulation of pathogenic streptococci. Methods Enzymol. 1991;204:556–586. doi: 10.1016/0076-6879(91)04028-m. [DOI] [PubMed] [Google Scholar]
  6. Caparon M. G., Scott J. R. Identification of a gene that regulates expression of M protein, the major virulence determinant of group A streptococci. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8677–8681. doi: 10.1073/pnas.84.23.8677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen C., Bormann N., Cleary P. P. VirR and Mry are homologous trans-acting regulators of M protein and C5a peptidase expression in group A streptococci. Mol Gen Genet. 1993 Dec;241(5-6):685–693. doi: 10.1007/BF00279912. [DOI] [PubMed] [Google Scholar]
  8. Chen L., Helmann J. D. Bacillus subtilis MrgA is a Dps(PexB) homologue: evidence for metalloregulation of an oxidative-stress gene. Mol Microbiol. 1995 Oct;18(2):295–300. doi: 10.1111/j.1365-2958.1995.mmi_18020295.x. [DOI] [PubMed] [Google Scholar]
  9. Chen L., Keramati L., Helmann J. D. Coordinate regulation of Bacillus subtilis peroxide stress genes by hydrogen peroxide and metal ions. Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8190–8194. doi: 10.1073/pnas.92.18.8190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Claiborne A., Ross R. P., Parsonage D. Flavin-linked peroxide reductases: protein-sulfenic acids and the oxidative stress response. Trends Biochem Sci. 1992 May;17(5):183–186. doi: 10.1016/0968-0004(92)90263-9. [DOI] [PubMed] [Google Scholar]
  11. Cleary P. P., Larkin A. Hyaluronic acid capsule: strategy for oxygen resistance in group A streptococci. J Bacteriol. 1979 Dec;140(3):1090–1097. doi: 10.1128/jb.140.3.1090-1097.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. DiRita V. J., Mekalanos J. J. Genetic regulation of bacterial virulence. Annu Rev Genet. 1989;23:455–482. doi: 10.1146/annurev.ge.23.120189.002323. [DOI] [PubMed] [Google Scholar]
  13. Dowds B. C. The oxidative stress response in Bacillus subtilis. FEMS Microbiol Lett. 1994 Dec 15;124(3):255–263. doi: 10.1111/j.1574-6968.1994.tb07294.x. [DOI] [PubMed] [Google Scholar]
  14. Eaton T., Shearman C., Gasson M. Cloning and sequence analysis of the dnaK gene region of Lactococcus lactis subsp. lactis. J Gen Microbiol. 1993 Dec;139(12):3253–3264. doi: 10.1099/00221287-139-12-3253. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Fogg G. C., Gibson C. M., Caparon M. G. The identification of rofA, a positive-acting regulatory component of prtF expression: use of an m gamma delta-based shuttle mutagenesis strategy in Streptococcus pyogenes. Mol Microbiol. 1994 Feb;11(4):671–684. doi: 10.1111/j.1365-2958.1994.tb00345.x. [DOI] [PubMed] [Google Scholar]
  17. Hanski E., Caparon M. Protein F, a fibronectin-binding protein, is an adhesin of the group A streptococcus Streptococcus pyogenes. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6172–6176. doi: 10.1073/pnas.89.13.6172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hanski E., Horwitz P. A., Caparon M. G. Expression of protein F, the fibronectin-binding protein of Streptococcus pyogenes JRS4, in heterologous streptococcal and enterococcal strains promotes their adherence to respiratory epithelial cells. Infect Immun. 1992 Dec;60(12):5119–5125. doi: 10.1128/iai.60.12.5119-5125.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lee J. Y., Caparon M. An oxygen-induced but protein F-independent fibronectin-binding pathway in Streptococcus pyogenes. Infect Immun. 1996 Feb;64(2):413–421. doi: 10.1128/iai.64.2.413-421.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Liochev S. I., Fridovich I. Fumarase C, the stable fumarase of Escherichia coli, is controlled by the soxRS regulon. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):5892–5896. doi: 10.1073/pnas.89.13.5892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Miller J. F., Mekalanos J. J., Falkow S. Coordinate regulation and sensory transduction in the control of bacterial virulence. Science. 1989 Feb 17;243(4893):916–922. doi: 10.1126/science.2537530. [DOI] [PubMed] [Google Scholar]
  22. Nakayama K. Nucleotide sequence of Streptococcus mutans superoxide dismutase gene and isolation of insertion mutants. J Bacteriol. 1992 Aug;174(15):4928–4934. doi: 10.1128/jb.174.15.4928-4934.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Okada N., Geist R. T., Caparon M. G. Positive transcriptional control of mry regulates virulence in the group A streptococcus. Mol Microbiol. 1993 Mar;7(6):893–903. doi: 10.1111/j.1365-2958.1993.tb01180.x. [DOI] [PubMed] [Google Scholar]
  24. Parker M. W., Blake C. C. Iron- and manganese-containing superoxide dismutases can be distinguished by analysis of their primary structures. FEBS Lett. 1988 Mar 14;229(2):377–382. doi: 10.1016/0014-5793(88)81160-8. [DOI] [PubMed] [Google Scholar]
  25. Perez-Casal J., Caparon M. G., Scott J. R. Mry, a trans-acting positive regulator of the M protein gene of Streptococcus pyogenes with similarity to the receptor proteins of two-component regulatory systems. J Bacteriol. 1991 Apr;173(8):2617–2624. doi: 10.1128/jb.173.8.2617-2624.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Podbielski A., Peterson J. A., Cleary P. Surface protein-CAT reporter fusions demonstrate differential gene expression in the vir regulon of Streptococcus pyogenes. Mol Microbiol. 1992 Aug;6(16):2253–2265. doi: 10.1111/j.1365-2958.1992.tb01401.x. [DOI] [PubMed] [Google Scholar]
  27. Privalle C. T., Fridovich I. Transcriptional and maturational effects of manganese and iron on the biosynthesis of manganese-superoxide dismutase in Escherichia coli. J Biol Chem. 1992 May 5;267(13):9140–9145. [PubMed] [Google Scholar]
  28. Ross R. P., Claiborne A. Cloning, sequence and overexpression of NADH peroxidase from Streptococcus faecalis 10C1. Structural relationship with the flavoprotein disulfide reductases. J Mol Biol. 1991 Oct 5;221(3):857–871. doi: 10.1016/0022-2836(91)80180-3. [DOI] [PubMed] [Google Scholar]
  29. Ross R. P., Claiborne A. Molecular cloning and analysis of the gene encoding the NADH oxidase from Streptococcus faecalis 10C1. Comparison with NADH peroxidase and the flavoprotein disulfide reductases. J Mol Biol. 1992 Oct 5;227(3):658–671. doi: 10.1016/0022-2836(92)90215-6. [DOI] [PubMed] [Google Scholar]
  30. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Schrum L. W., Hassan H. M. Transcriptional activation of Mn-superoxide dismutase gene (sodA) of Escherichia coli by MnCl2. Biochim Biophys Acta. 1993 Nov 16;1216(2):186–190. doi: 10.1016/0167-4781(93)90143-2. [DOI] [PubMed] [Google Scholar]
  32. Scott J. R. A turbid plaque-forming mutant of phage P1 that cannot lysogenize Escherichia coli. Virology. 1974 Dec;62(2):344–349. doi: 10.1016/0042-6822(74)90397-3. [DOI] [PubMed] [Google Scholar]
  33. Scott J. R., Guenthner P. C., Malone L. M., Fischetti V. A. Conversion of an M- group A streptococcus to M+ by transfer of a plasmid containing an M6 gene. J Exp Med. 1986 Nov 1;164(5):1641–1651. doi: 10.1084/jem.164.5.1641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shaw W. V. Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria. Methods Enzymol. 1975;43:737–755. doi: 10.1016/0076-6879(75)43141-x. [DOI] [PubMed] [Google Scholar]
  35. Simpson W. J., LaPenta D., Chen C., Cleary P. P. Coregulation of type 12 M protein and streptococcal C5a peptidase genes in group A streptococci: evidence for a virulence regulon controlled by the virR locus. J Bacteriol. 1990 Feb;172(2):696–700. doi: 10.1128/jb.172.2.696-700.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  38. Stevens D. L. Invasive group A streptococcus infections. Clin Infect Dis. 1992 Jan;14(1):2–11. doi: 10.1093/clinids/14.1.2. [DOI] [PubMed] [Google Scholar]
  39. VanHeyningen T., Fogg G., Yates D., Hanski E., Caparon M. Adherence and fibronectin binding are environmentally regulated in the group A streptococci. Mol Microbiol. 1993 Sep;9(6):1213–1222. doi: 10.1111/j.1365-2958.1993.tb01250.x. [DOI] [PubMed] [Google Scholar]
  40. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  41. Voelker U., Voelker A., Maul B., Hecker M., Dufour A., Haldenwang W. G. Separate mechanisms activate sigma B of Bacillus subtilis in response to environmental and metabolic stresses. J Bacteriol. 1995 Jul;177(13):3771–3780. doi: 10.1128/jb.177.13.3771-3780.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Völker U., Engelmann S., Maul B., Riethdorf S., Völker A., Schmid R., Mach H., Hecker M. Analysis of the induction of general stress proteins of Bacillus subtilis. Microbiology. 1994 Apr;140(Pt 4):741–752. doi: 10.1099/00221287-140-4-741. [DOI] [PubMed] [Google Scholar]
  43. Wannamaker L. W. Differences between streptococcal infections of the throat and of the skin. I. N Engl J Med. 1970 Jan 1;282(1):23–31. doi: 10.1056/NEJM197001012820106. [DOI] [PubMed] [Google Scholar]
  44. Wu J., Weiss B. Two divergently transcribed genes, soxR and soxS, control a superoxide response regulon of Escherichia coli. J Bacteriol. 1991 May;173(9):2864–2871. doi: 10.1128/jb.173.9.2864-2871.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES