Antonie van Leeuwenhoek

, Volume 51, Issue 2, pp 203–218 | Cite as

Plasmid-determined cloacin DF13-susceptibility inEnterobacter cloacae andKlebsiella edwardsii; identification of the cloacin DF13/aerobactin outer membrane receptor proteins

  • Willy J. A. Krone
  • Gregory Koningstein
  • Frits K. de Graaf
  • Bauke Oudega
Genetics and Molecular Biology


BothEnterobacter cloacae H478 andKlebsiella edwardsii S15 were shown to harbour a relatively large conjugative plasmid that coded for cloacin DF13-susceptibility and the production and uptake of a hydroxamate iron chelator, most probably aerobactin. Protein-blotting experiments with antiserum raised against the purified cloacin DF13/aerobactin receptor protein fromEscherichia coli (Co1V-K30) revealed that the corresponding outer membrane receptor proteins ofEnt. cloacae H478 andK. edwardsii S15 had apparent mol wts of 85 000 and 76000, respectively.E. coli transconjugants harbouring either the plasmid fromEnt. cloacae H478 orK. edwardsii S15 expressed a cloacin DF13/aerobactin outer membrane receptor protein with a mol wt of 74000. The receptor protein encoded by theEnt. cloacae andK. edwardsii plasmids were immunologically more related to each other than to the pCo1V-K30-encoded receptor protein.


Iron Outer Membrane Receptor Protein Membrane Receptor Iron Chelator 
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  1. Bolivar, F., Rodriguez, R. L., Greene, P. J., Betlach, M. C., HEyneker, H. L., Boyer, H. W., Crosa, J. H. andFalkow, S. 1977. Construction and characterization of new cloning vehicles 11. A multipurpose cloning system. -Gene2: 95–113.CrossRefPubMedGoogle Scholar
  2. Bowles, L. K. andKonisky, J. 1981. Cleavage of colicin la by theEscherichia coli K-12 outer membrane is not mediated by the colicin Ia receptor. -J. Bacteriol.145: 668–671.PubMedGoogle Scholar
  3. Braun, V. 1981.Escherichia coli cells containing the plasmid ColV produce the iron ionophore aerobactin. -FEMS Microbiol. Lett.11: 225–228.CrossRefGoogle Scholar
  4. Braun, V., Gross, R., Koster, W. andZimmermann, L. 1983. Plasmid and chromosomal mutants in the iron (III)-aerobactin transport system ofEscherichia coli. Use of streptonigrin for selection. -Mol. Gen. Genet.192: 131–139.CrossRefPubMedGoogle Scholar
  5. Chang, A. C. Y. andCohen, S. N. 1978. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P 15A cryptic miniplasmid. -J. Bacteriol.134: 1141–1156.PubMedGoogle Scholar
  6. Cohen, S. N., Chang, A. C. Y. andHsu, L. 1972. Nonchromosomal antibiotic resistance in bacteria: genetic transformation ofEscherichia coli by R-factor DNA. -Proc. Natl Acad. Sci. USA69: 2110–2114.PubMedGoogle Scholar
  7. Csaky, T. Z. 1948. On the estimation of bound hydroxylamine in biological materials. -Acta Chem. Scand.2: 450–454.Google Scholar
  8. De Graaf, F. K. andKlaasen-Boor, P. 1977. Purification and characterization of a complex between cloacin and its immunity protein isolated fromEnterobacter cloacae (Clo DF13). -Eur. J. Biochem.73: 107–114.CrossRefPubMedGoogle Scholar
  9. De Graaf, F. K., Niekus, H. G. D. andKlootwijk, J. 1973. Inactivation of bacterial ribosomes in vivo and in vitro by cloacin DF13. -FEBS Lett.35: 161–165.CrossRefPubMedGoogle Scholar
  10. De Graaf, F. K., Spanjaerdt Speckman, E. A. andStouthamer, A. H. 1969. Mode of action of a bacteriôcin produced byEnterobacter cloacae DF13. -Antonie van Leeuwenhoek35: 287–306.CrossRefPubMedGoogle Scholar
  11. Fiss, E. H., Hollifield Jr, W. C. andNeilands, J. B. 1979. Absence of ferric enterobactin receptor modification activity in mutants ofEscherichia coli K-12 lacking proteina. -Biochem. Biophys. Res. Commun.91: 29–34.CrossRefPubMedGoogle Scholar
  12. Fiss, E. H., Stanley-Samuelson, P. andNeilands, J. B. 1982. Properties and proteolysis of ferric enterobactin outer membrane receptor inEscherichia coli K12. -Biochemistry21: 4517–4522.CrossRefPubMedGoogle Scholar
  13. Grewal, K. K., Warner, P. J. andWilliams, P. H. 1982. An inducible outer membrane protein involved in aerobactin-mediated iron transport by Co1V strains ofEscherichia coli. -FEBS Lett.140: 27–30.CrossRefPubMedGoogle Scholar
  14. Gross, R., Engelsrecht, F. andBraun, V. 1984. Genetic and biochemical characterization of the aerobactin synthesis operon on pCo1V. -Mol. Gen. Genet.196: 74–80.CrossRefPubMedGoogle Scholar
  15. Hawkes, R., Niday, E. andGordon, J. 1982. A dot-immunobinding assay for monoclonal and other antibodies. -Anal. Biochem.119: 142–147.CrossRefPubMedGoogle Scholar
  16. Krone, W. J. A., Luirink, J., Koningstein, G., Oudega, B. andDe Graaf, F. K. 1983a. Subcloning of the cloacin DF13/aerobactin receptor protein and identification of a pCo1V-K30-determined polypeptide involved in ferric-aerobactin uptake. -J. Bacteriol.156: 945–948.PubMedGoogle Scholar
  17. Krone, W. J. A., Oudega, B., Stegehuis, F. andDe Graaf, F. K. 1983b. Cloning and expression of the cloacin DF13/aerobactin receptor ofEscherichia coli (Co1V-K30). -J. Bacteriol.153: 716–721.PubMedGoogle Scholar
  18. Krone, W. J. A., Stegehuis, F., Koningstein, G., Van Doorn, C., Roosendaal, B., De Graaf, F. K. andOudega, B. 1985. Characterization of the pCo l V-K30 encoded cloacin DF 13/aerobactin outer membrane receptor protein ofEscherichia coli; isolation and purification of the protein and analysis of its nucleotide sequence and primary structure. -FEMS Microbiol. Lett.26: 153–161.CrossRefGoogle Scholar
  19. Lawlor, K. M. andPayne, S. M. 1984. Aerobactin genes inShigella spp. -J. Bacteriol.160: 266–272.PubMedGoogle Scholar
  20. Lugtenberg, B., Meijers, J., Peters, R., Van der Hoek, P. andVan Alpren, L. 1975. Electrophoretic resolution of the ‘major outer membrane protein’ ofEscherichia coli K12 into four bands. -FEBS Lett.58: 254–258.CrossRefPubMedGoogle Scholar
  21. Maniatis, T., Fritsch, E. F. andSambrook, J. 1982. Molecular Cloning. -Cold Spring Harbor, New York.Google Scholar
  22. McDougall, S. andNeilands, J. B. 1984. Plasmid- and chromosome-coded aerobactin synthesis in enteric bacteria: insertion sequences flank operon in plasmid-mediated systems. -J. Bacteriol.159: 300–305.PubMedGoogle Scholar
  23. Oudega, B., Oldenziel-Werner, W. J. M., Klaasen-Boor, P., Rezee, A., Glas, J. andDe Graaf, F. K. 1979. Purification and characterization of cloacin DF13 receptor fromEnterobacter cloacae and its interaction with cloacin DF13 in vitro. -J. Bacteriol.138: 7–16.PubMedGoogle Scholar
  24. Perez-Casal, J. F. andCrosa, J. H. 1984. Aerobactin iron uptake sequences in plasmid Co1V-K30 are flanked by inverted IS1-like elements and replication regions. -J. Bacteriol.160: 256–265.PubMedGoogle Scholar
  25. Prakash, R. K., Schilperoort, R. A. andNuti, M. P. 1981. Large plasmids of fast-growing rhizobia: homology studies and location of structural nitrogen fixation (nif) genes. -J. Bacteriol.145: 1129–1136.PubMedGoogle Scholar
  26. Régnier, P. 1981. Identification of protease IV ofE. coli: an outer membrane bound enzyme. -Biochem. Biophys. Res. Commun.99: 844–854.PubMedGoogle Scholar
  27. Sato, T. andYura, T. 1979. Chromosomal location and expression of the structural gene for major outer membrane protein Ia ofEscherichia coli K-12 and of the homologous gene of Salmonella typhimurium. -J. Bacteriol.139: 468–477.PubMedGoogle Scholar
  28. Stouthamer, A. H. andTieze, G. A. 1966. Bacteriocin production by members of the genusKlebsiella. -Antonie van Leeuwenhock32: 171–182.Google Scholar
  29. Stuart, S. J., Greenwood, K. T. andLuke, R. K. J. 1980. Hydroxamate-mediated transport of iron controlled by Co1V plasmids. -J. Bacteriol.143: 35–42.PubMedGoogle Scholar
  30. Towbin, H., Staehelin, T. andGordon, J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. -Proc. Natl Acad. Sci. USA76: 4350–4354.PubMedGoogle Scholar
  31. Valvano, M. A. andCrosa, J. H. 1984. Aerobactin iron transport genes commonly encoded by certain Co1V plasmids occur in the chromosome of a human invasive strain ofEscherichia coli Kl. -Infect. Immun.46: 159–167.PubMedGoogle Scholar
  32. Van Tiel-Menkveld, G. J., Mentjox-Vervuurt, J. M., Oudega., B. andDe Graaf, F. K. 1982. Siderophore production byEnterobacter cloacae and a common receptor protein for the uptake of aerobactin and cloacin DF13. -J. Bacteriol.150: 490–497.PubMedGoogle Scholar
  33. Van Tiel-Menkveld, G. J., Oudega, B., Kempers, O. andDe Graaf, F. K. 1981. The possible involvement of the cloacin DF13 receptor protein in the hydroxamate-mediated uptake of iron byEnterobacter cloacae andEscherichia coli (Co1V). -FEMS Microbiol. Lett.12: 373–380.Google Scholar
  34. Van Tiel-Menkveld, G. J., Rezef, A. andDe Graaf, F. K. 1979. Production and excretion of cloacin DF13 byEscherichia coli harboring plasmid CloDF13. -J. Bacteriol.140: 415–423.Google Scholar
  35. Wagegg, W. andBraun, V. 1981. Ferric citrate transport inEscherichia coli requires outer membrane receptor protein FecA. -J. Bacteriol.145: 156–163.PubMedGoogle Scholar
  36. Warner, P. J., Williams, P. H., Bindereif, A. andNeilands, J. B. 1981. ColV plasmid-specified aerobactin synthesis by invasive strains ofEscherichia coli. -Infect. Immun.33: 540–545.PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1985

Authors and Affiliations

  • Willy J. A. Krone
    • 1
  • Gregory Koningstein
    • 1
  • Frits K. de Graaf
    • 1
  • Bauke Oudega
    • 1
  1. 1.Department of Microbiology, Biological LaboratoryVrije UniversiteitAmsterdamThe Netherlands

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