Molecular and General Genetics MGG

, Volume 210, Issue 1, pp 135–139 | Cite as

Selection procedure for deregulated iron transport mutants (fur) in Escherichia coli K 12: fur not only affects iron metabolism

  • Klaus Hantke


A selection procedure using Mn2+ is described. A high percentage of the Mn2+ resistant mutants had constitutive iron transport systems. By P1 transduction, and complementation with the cloned fur gene it could be shown that nearly all the mutants constitutive in the expression of the operon fusion fiu::λplacMu were only defective in fur. High concentrations of manganese inhibited the derepression of an iron-regulated lac operon fusion. In another iron-regulated lac operon fusion that was inducible by iron, manganese also induced the production of β-galactosidase. Most of the fur mutants isolated (80%) were not able to grow on succinate, fumarate or acetate. After transformation with a fur+ plasmid all 39 mutants tested were able to grow on succinate. In fur mutants the presence of succinate in the growth medium reduced succinate uptake rates by 50%–70%. Succinate dehydrogenase activity was reduced to 10% of that of the parent strain.

Key words

fur selection with Mn2+ Mn2+ and iron transport Succinate utilization 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Anwar H, Brown MRW, Day A, Weller PH (1984) Outer membrane antigens of mucoid Pseudomonas aeruginosa isolated directly from the sputum of a cystic fibrosis patient. FEMS Microbiol Lett 24:235–239Google Scholar
  2. Bewick MA, Lo TCY (1979) Dicarboxylic acid transport in Escherichia coli K12: involvement of a binding protein in the translocation of dicarboxylic acids across the outer membrane of the cell envelope. Can J Biochem 57:653–661Google Scholar
  3. Bremer E, Silhavy TS, Weinstock GM (1985) Transposable λplacMu bacteriophages for creating lacZ operon fusions and kanamycin resistance insertions in Escherichia coli. J Bacteriol 162:1092–1099Google Scholar
  4. Fiedler W, Rotering H (1985) Characterization of an Escherichia coli mdoB mutant strain unable to transfer sn-1-phosphoglycerol to membrane-derived-oligosaccharides. J Biol Chem 260:4799–4806Google Scholar
  5. Hantke K (1981) Regulation of ferric iron transport in Escherichia coli K12: isolation of a constitutive mutant. Mol Gen Genet 182:288–292Google Scholar
  6. Hantke K (1983) Identification of an iron uptake system specific for coprogen and rhodotorulic acid in Escherichia coli. Mol Gen Genet 191:301–306Google Scholar
  7. Hantke K (1984) Cloning of the repressor protein gene of iron regulated systems in Escherichia coli K12. Mol Gen Genet 197:337–341Google Scholar
  8. Hantke K (1985) Iron transport in bacteria. In: Spik G, Montreuil J, Crichton RR, Mazurier J (eds) Proteins of iron storage and transport in bacteria. Elsevier, Amsterdam, pp 231–243Google Scholar
  9. Hedblom ML, Adler J (1980) Genetic and biochemical properties of Escherichia coli mutants with defects in serine chemotaxis. J Bacteriol 144:1048–1060Google Scholar
  10. Herbert AA, Guest JR (1971) Two mutations affecting the utilization of C4-dicarboxylic acids by Escherichia coli. J Gen Microbiol 63:151–162Google Scholar
  11. Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  12. Montie TC, Doyle-Huntzinger D, Craven RC, Holder IA (1982) Loss of virulence associated with absence of flagellum in an isogenic mutant of Pseudomonas aeruginosa in the burned mouse model. Infect Immun 38:1296–1298Google Scholar
  13. Osborn MJ, Gander JE, Parisi E, Carson J (1972) Mechanism of assembly of the outer membrane of Salmonella typhimurium. J Biol Chem 247:3962–3972Google Scholar
  14. Rainnie DJ, Bragg PD (1973) The effect of iron deficiency on respiration and energy coupling in Escherichia coli. J Gen Microbiol 77:339–349Google Scholar
  15. Schäffer S, Hantke K, Braun V (1985) Nucleotide sequence of the regulatory gene fur. Mol Gen Genet 200:110–113Google Scholar
  16. Silver S, Johnseine P, Whitney E, Clark D (1972) Manganeseresistant mutants of Escherichia coli: Physiological and genetic studies. J Bacteriol 110:186–195Google Scholar
  17. Vogel HJ, Bonner DM (1956) A convenient growth medium for Escherichia coli and some other micro-organisms. Microbiol Genet Bull 13:43–44Google Scholar

Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • Klaus Hantke
    • 1
  1. 1.Lehrstuhl für Mikrobiologie IIUniversität TübingenTübingenFederal Republic of Germany

Personalised recommendations