Advertisement

Molecular and General Genetics MGG

, Volume 196, Issue 1, pp 74–80 | Cite as

Genetic and biochemical characterization of the aerobactin synthesis operon on pColV

  • Roy Gross
  • Fred Engelbrecht
  • Volkmar Braun
Article

Summary

Certain ColV plasmids of Escherichia coli contain genes that specify functions for the acquisition of iron-(III) via aerobactin. The locus for aerobactin synthesis of pColV-K311 was cloned into pBR322. Mutagenesis with the transposon Tn1000, and the generation of deletions with restriction enzymes resulted in multicopy plasmids which complemented pColV mutants impaired in various steps of aerobactin synthesis. The insertion and deletion mutants were mapped and assigned to three loci termed aerA, aerB, and aerC. It is proposed that these genes mediate the synthesis of aerobactin by specifying functions for hydroxylation (aer A), acetylation of the 6-amino group of 6-hydroxylysine (aer B), and the coupling of N-acetyl-N-hydroxy-lysine with citrate (aerC). The order and transcription polarity of the structural genes was found to be aer B, aerC, aer A.

Keywords

Enzyme Iron Escherichia Coli Citrate Restriction Enzyme 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Aisen P, Leibman A (1972) Lactoferrin and transferrin: a comparative study. Biochim Biophys Acta 257:314–323Google Scholar
  2. Archibald F (1983) Lactobacillus plantarum, an organism not requiring iron. FEMS Microbiol Lett 19:29–32Google Scholar
  3. Bindereif A, Neilands JB (1983) Cloning of the aerobactin-mediated iron assimilation system of plasmid ColV. J Bacteriol 153:1111–1113Google Scholar
  4. Bindereif A, Braun V, Hantke K (1982) The cloacin receptor of ColV-bearing Escherichia coli is part of the Fe3+-aerobactin transport system. J Bacteriol 150:1472–1475Google Scholar
  5. Birnboim HC (1983) A rapid alkaline extraction method for the isolation of plasmid DNA. In: Wu R, Grossman L, Moldave K (eds) Methods in enzymology 100. Academic Press, New York, pp 243–255Google Scholar
  6. Bolivar F (1978) Derivatives of plasmid pBR322 in genetic engineering. In: Boyer HW, Nicosia S (eds) Molecular cloning vectors. Elsevier, North Holland Biomedical Press, Amsterdam, New York, pp 95–163Google Scholar
  7. Braun V (1981) Escherichia coli containing the plasmid ColV produce the iron ionophore aerobactin. FEMS Microbiol Lett 11:215–228Google Scholar
  8. Braun V, Burkhardt R (1982) Regulation of the ColV plasmid-determined iron(III)-aerobactin transport system in Escherichia coli. J Bacteriol 152:223–231Google Scholar
  9. Braun V, Burkhardt R, Schneider R, Zimmermann L (1982) Chromosomal genes for ColV plasmid-determined Fe3+-aerobactin transport in Escherichia coli. J Bacteriol 152:553–559Google Scholar
  10. Braun V, Gross R, Köster W, Zimmermann L (1983) Plasmid and chromosomal mutants in the iron(III)-aerobactin transport system of Escherichia coli. Use of streptonigrin for selection. Mol Gen Genet 192:131–139Google Scholar
  11. Braun V, Brazel-Faisst C, Schneider R (1984) Growth stimulation of Escherichia coli in serum by iron(III) aerobactin. Recycling of aerobactin. FEMS Microbiol Lett 21:99–103Google Scholar
  12. Csáky T (1948) On the estimation of bound hydroxylamine in biological materials. Acta Chem Scand 2:450–454Google Scholar
  13. Fecker L, Braun V (1983) Cloning and expression of the fhu genes involved in the iron(III)-hydroxamate uptake of Escherichia coli. J Bacteriol 156:1301–1314Google Scholar
  14. Gibson F, Magrath DI (1969) The isolation and characterization of a hydroxamic acid (aerobactin) formed by Aerobacter aerogenes 62-1. Biochim Biophys Acta 192:175–184Google Scholar
  15. Grewal KK, Warner PJ, Williams PH (1982) An inducible outer membrane protein involved in aerobactin-mediated iron-transport by ColV strains of Escherichia coli. FEBS Lett 140:27–30Google Scholar
  16. Guyer MS (1983) Uses of transposon γδ in the analysis of cloned genes. In: Wu R, Grossman L, Moldave K (eds) Methods in enzymology 101. Academic Press, New York, pp 362–369Google Scholar
  17. Krone WJA, Oudega B, Stegehuis F, de Graaf F (1983a) Cloning and expression of the cloacin DF13/aerobactin receptor of Escherichia coli (ColV-K30). J Bacteriol 153:716–721Google Scholar
  18. Krone WJA, Luirink J, Koningstein G, Oudega B, de Graaf F (1983b) Subcloning of the cloacin DF13/aerobactin receptor protein and identification of a pColV-K30-determined polypeptide involved in ferric-aerobactin uptake. J Bacteriol 156:945–948Google Scholar
  19. Maurer PJ, Miller MJ (1982) Microbial iron chelators: total synthesis of aerobactin and its constituent amino acid, N6-acetyl-N6-hydroxylysine. J Am Chem Soc 104:3096–3101Google Scholar
  20. Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  21. Monod J, Cohen-Bazire G, Cohn M (1951) Sur la biosynthése de la β-galactosidase (lactase) chez Escherichia coli. La spécificité de l'induction. Biochim Biophys Acta 7:585–599Google Scholar
  22. Murray GJ, Clark GED, Parniak MA, Viswanatha T (1977) Effect of metabolites on ɛ-N-hydroxylysine formation in cell-free extracts of Aerobacter aerogenes 62-1. Can J Biochem 55:625–629Google Scholar
  23. Parniak MA, Jackson GED, Murray GJ, Viswanatha T (1979) Studies on the formation of N6-hydroxylysine in cell-free extracts of Aerobacter aerogenes 62-1. Biochim Biophys Acta 569:99–108Google Scholar
  24. van Tiel-Menkveld GJ, Oudega GJB, Kempers O, de Graaf FK (1981) The possible involvement of the cloacin DF13 receptor protein in the hydroxamate-mediated uptake of iron by Enterobacter cloacae and Escherichia coli (ColV). FEMS Microbiol Lett 12:373–380Google Scholar
  25. Tomlinson G, Cruickshank WH, Viswanatha T (1971) Sensitivity of substituted hydroxylamines to determination by iodine oxidation. Anal Biochem 44:670–679Google Scholar
  26. Warner PJ, Williams PH, Bindereif A, Neilands JB (1981) ColV plasmid-specified aerobactin synthesis by invasive strains of Escherichia coli. Infect Immun 33:540–545Google Scholar
  27. Weinberg ED (1978) Iron and infection. Microbiol Rev 42:45–66Google Scholar
  28. Williams PH (1979) Novel iron uptake system specified by ColV plasmids: an important component in the virulence of invasive strains of Escherichia coli. Infect Immun 26:925–932Google Scholar
  29. Williams PH, Warner PJ (1980) ColV plasmid-mediated, colicin V-independent iron uptake system of invasive strains of Escherichia coli. Infect Immun 29:411–416Google Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • Roy Gross
    • 1
  • Fred Engelbrecht
    • 1
  • Volkmar Braun
    • 1
  1. 1.Mikrobiologie Il der UniversitätTübingenFederal Republic of Germany

Personalised recommendations