Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1989 Sep;57(9):2794–2798. doi: 10.1128/iai.57.9.2794-2798.1989

Expression in Escherichia coli K-12 of the 76,000-dalton iron-regulated outer membrane protein of Shigella flexneri confers sensitivity to cloacin DF13 in the absence of Shigella O antigen.

P Derbyshire 1, T Baldwin 1, P Stevenson 1, E Griffiths 1, M Roberts 1, P Williams 1, T L Hale 1, S B Formal 1
PMCID: PMC313528  PMID: 2474501

Abstract

One of the chromosomal segments associated with virulence in Shigella flexneri encodes the production of aerobactin and the synthesis of an iron-regulated 76-kilodalton outer membrane protein believed to be the ferric-aerobactin receptor. However, S. flexneri expressing this putative aerobactin receptor, which is slightly larger than that encoded by pColV, is insensitive to the killing action of cloacin DF13, a bacteriocin which binds to other aerobactin receptor proteins and kills the cells. In this paper we show that the conjugal transfer of DNA encoding the iron-regulated 76-kilodalton protein from S. flexneri to Escherichia coli K-12 conferred cloacin DF13 sensitivity on the recipients. However, E. coli K-12 which had also inherited genes specifying Shigella O-antigen biosynthesis remained cloacin insensitive. The data suggest that it is unwise to use cloacin DF13 sensitivity alone to screen transconjugants or clinical isolates for the expression of aerobactin receptor proteins.

Full text

PDF
2794

Images in this article

2796Image
on p.2796
2797Image
on p.2797

Selected References

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

  1. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bindereif A., Neilands J. B. Cloning of the aerobactin-mediated iron assimilation system of plasmid ColV. J Bacteriol. 1983 Feb;153(2):1111–1113. doi: 10.1128/jb.153.2.1111-1113.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bywater M., Bywater R., Hellman L. A novel chromatographic procedure for purification of bacterial plasmids. Anal Biochem. 1983 Jul 1;132(1):219–224. doi: 10.1016/0003-2697(83)90451-7. [DOI] [PubMed] [Google Scholar]
  4. Chart H., Griffiths E. Antigenic and molecular homology of the ferric enterobactin receptor protein of Escherichia coli. J Gen Microbiol. 1985 Jun;131(6):1503–1509. doi: 10.1099/00221287-131-6-1503. [DOI] [PubMed] [Google Scholar]
  5. Chart H., Shaw D. H., Ishiguro E. E., Trust T. J. Structural and immunochemical homogeneity of Aeromonas salmonicida lipopolysaccharide. J Bacteriol. 1984 Apr;158(1):16–22. doi: 10.1128/jb.158.1.16-22.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. De Graaf F. K., Klaasen-Boor P. Purification and characterization of a complex between cloacin and its immunity protein isolated from Enterobacter cloacae (Clo DF13). Dissociation and reconstitution of the complex. Eur J Biochem. 1977 Feb 15;73(1):107–114. doi: 10.1111/j.1432-1033.1977.tb11296.x. [DOI] [PubMed] [Google Scholar]
  7. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  8. Formal S. B., Gemski P., Baron L. S., Labrec E. H. Genetic Transfer of Shigella flexneri Antigens to Escherichia coli K-12. Infect Immun. 1970 Mar;1(3):279–287. doi: 10.1128/iai.1.3.279-287.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Griffiths E., Stevenson P., Hale T. L., Formal S. B. Synthesis of aerobactin and a 76,000-dalton iron-regulated outer membrane protein by Escherichia coli K-12-Shigella flexneri hybrids and by enteroinvasive strains of Escherichia coli. Infect Immun. 1985 Jul;49(1):67–71. doi: 10.1128/iai.49.1.67-71.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gross R., Engelbrecht F., Braun V. Identification of the genes and their polypeptide products responsible for aerobactin synthesis by pColV plasmids. Mol Gen Genet. 1985;201(2):204–212. doi: 10.1007/BF00425661. [DOI] [PubMed] [Google Scholar]
  11. Hale T. L., Formal S. B. Genetics of virulence in Shigella. Microb Pathog. 1986 Dec;1(6):511–518. doi: 10.1016/0882-4010(86)90037-9. [DOI] [PubMed] [Google Scholar]
  12. Hale T. L., Sansonetti P. J., Schad P. A., Austin S., Formal S. B. Characterization of virulence plasmids and plasmid-associated outer membrane proteins in Shigella flexneri, Shigella sonnei, and Escherichia coli. Infect Immun. 1983 Apr;40(1):340–350. doi: 10.1128/iai.40.1.340-350.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hollifield W. C., Jr, Fiss E. H., Neilands J. B. Modification of a ferric enterobactin receptor protein from the outer membrane of Escherichia coli. Biochem Biophys Res Commun. 1978 Jul 28;83(2):739–746. doi: 10.1016/0006-291x(78)91051-3. [DOI] [PubMed] [Google Scholar]
  14. Lafont J. P., Dho M., D'Hauteville H. M., Bree A., Sansonetti P. J. Presence and expression of aerobactin genes in virulent avian strains of Escherichia coli. Infect Immun. 1987 Jan;55(1):193–197. doi: 10.1128/iai.55.1.193-197.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lawlor K. M., Daskaleros P. A., Robinson R. E., Payne S. M. Virulence of iron transport mutants of Shigella flexneri and utilization of host iron compounds. Infect Immun. 1987 Mar;55(3):594–599. doi: 10.1128/iai.55.3.594-599.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Linggood M. A., Roberts M., Ford S., Parry S. H., Williams P. H. Incidence of the aerobactin iron uptake system among Escherichia coli isolates from infections of farm animals. J Gen Microbiol. 1987 Apr;133(4):835–842. doi: 10.1099/00221287-133-4-835. [DOI] [PubMed] [Google Scholar]
  17. MONOD J., COHEN-BAZIRE G., COHN M. Sur la biosynthèse de la beta-galactosidase (lactase) chez Escherichia coli; la spécificité de l'induction. Biochim Biophys Acta. 1951 Nov;7(4):585–599. doi: 10.1016/0006-3002(51)90072-8. [DOI] [PubMed] [Google Scholar]
  18. Marolda C. L., Valvano M. A., Lawlor K. M., Payne S. M., Crosa J. H. Flanking and internal regions of chromosomal genes mediating aerobactin iron uptake systems in enteroinvasive Escherichia coli and Shigella flexneri. J Gen Microbiol. 1987 Aug;133(8):2269–2278. doi: 10.1099/00221287-133-8-2269. [DOI] [PubMed] [Google Scholar]
  19. Nassif X., Mazert M. C., Mounier J., Sansonetti P. J. Evaluation with an iuc::Tn10 mutant of the role of aerobactin production in the virulence of Shigella flexneri. Infect Immun. 1987 Sep;55(9):1963–1969. doi: 10.1128/iai.55.9.1963-1969.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nassif X., Sansonetti P. J. Correlation of the virulence of Klebsiella pneumoniae K1 and K2 with the presence of a plasmid encoding aerobactin. Infect Immun. 1986 Dec;54(3):603–608. doi: 10.1128/iai.54.3.603-608.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Payne S. M., Niesel D. W., Peixotto S. S., Lawlor K. M. Expression of hydroxamate and phenolate siderophores by Shigella flexneri. J Bacteriol. 1983 Sep;155(3):949–955. doi: 10.1128/jb.155.3.949-955.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pugsley A. P. Escherichia coli K12 strains for use in the identification and characterization of colicins. J Gen Microbiol. 1985 Feb;131(2):369–376. doi: 10.1099/00221287-131-2-369. [DOI] [PubMed] [Google Scholar]
  23. Rogers H. J. Iron-Binding Catechols and Virulence in Escherichia coli. Infect Immun. 1973 Mar;7(3):445–456. doi: 10.1128/iai.7.3.445-456.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. SIMON E. H., TESSMAN I. THYMIDINE-REQUIRING MUTANTS OF PHAGE T4. Proc Natl Acad Sci U S A. 1963 Sep;50:526–532. doi: 10.1073/pnas.50.3.526. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sansonetti P. J., Hale T. L., Dammin G. J., Kapfer C., Collins H. H., Jr, Formal S. B. Alterations in the pathogenicity of Escherichia coli K-12 after transfer of plasmid and chromosomal genes from Shigella flexneri. Infect Immun. 1983 Mar;39(3):1392–1402. doi: 10.1128/iai.39.3.1392-1402.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Singh L., Jones K. W. The use of heparin as a simple cost-effective means of controlling background in nucleic acid hybridization procedures. Nucleic Acids Res. 1984 Jul 25;12(14):5627–5638. doi: 10.1093/nar/12.14.5627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  28. Van Tiel-Menkveld G. J., Mentjox-Vervuurt J. M., Oudega B., de Graaf F. K. Siderophore production by Enterobacter cloacae and a common receptor protein for the uptake of aerobactin and cloacin DF13. J Bacteriol. 1982 May;150(2):490–497. doi: 10.1128/jb.150.2.490-497.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wieslander L. A simple method to recover intact high molecular weight RNA and DNA after electrophoretic separation in low gelling temperature agarose gels. Anal Biochem. 1979 Oct 1;98(2):305–309. doi: 10.1016/0003-2697(79)90145-3. [DOI] [PubMed] [Google Scholar]
  30. Wray W., Boulikas T., Wray V. P., Hancock R. Silver staining of proteins in polyacrylamide gels. Anal Biochem. 1981 Nov 15;118(1):197–203. doi: 10.1016/0003-2697(81)90179-2. [DOI] [PubMed] [Google Scholar]
  31. de Graaf F. K., Spanjaerdt Speckman E. A., Stouthamer A. H. Mode of action of a bacteriocin produced by Enterobacter cloacae DF13. Antonie Van Leeuwenhoek. 1969;35(3):287–306. doi: 10.1007/BF02219150. [DOI] [PubMed] [Google Scholar]
  32. van der Ley P., de Graaff P., Tommassen J. Shielding of Escherichia coli outer membrane proteins as receptors for bacteriophages and colicins by O-antigenic chains of lipopolysaccharide. J Bacteriol. 1986 Oct;168(1):449–451. doi: 10.1128/jb.168.1.449-451.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES