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J Bacteriol. Jul 1987; 169(7): 3340–3349.
PMCID: PMC212388

Regulation of Escherichia coli fumarate reductase (frdABCD) operon expression by respiratory electron acceptors and the fnr gene product.

Abstract

The fumarate reductase enzyme complex, encoded by the frdABCD operon, allows Escherichia coli to utilize fumarate as a terminal electron acceptor for anaerobic oxidative phosphorylation. To analyze the expression of fumarate reductase, protein and operon fusions were constructed between the frdA and the lacZ genes and introduced onto the E. coli chromosome at the lambda attachment site. Expression of beta-galactosidase from either fusion was increased 10-fold during anaerobic versus aerobic cell growth, increased an additional 1.5-fold by the presence of fumarate, the substrate, and decreased 23-fold by nitrate, a preferred electron acceptor. The addition of trimethylamine-N-oxide as an electron acceptor did not significantly alter frdA'-'lacZ expression. Control of frd operon expression is therefore exerted at the transcriptional level in response to the availability of the electron acceptors oxygen, fumarate, and nitrate. Anaerobic induction of frdA'-'lacZ expression was impaired in an fnr mutant and was restored when the fnr+ gene was provided in trans, thus establishing that the fnr gene product, Fnr, is responsible for the anaerobic activation of frd operon expression. Nitrate repression of frdA'-'lacZ expression was observed under either aerobic or anaerobic cell growth conditions in both wild-type and fnr mutant strains, demonstrating that the mechanism for nitrate repression is independent of nitrate respiration and oxygen control imparted by Fnr. Studies performed with a fnr'-'lacZ protein fusion confirmed that the fnr gene is expressed both aerobically and anaerobically. A model is proposed for the regulation of frdABCD operon expression in response to the availability of the alternate terminal electron acceptors oxygen, nitrate, and fumarate.

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Selected References

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  • Ausubel FM. Regulation of nitrogen fixation genes. Cell. 1984 May;37(1):5–6. [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Casadaban MJ, Chou J, Cohen SN. In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals. J Bacteriol. 1980 Aug;143(2):971–980. [PMC free article] [PubMed]
  • Cecchini G, Ackrell BA, Deshler JO, Gunsalus RP. Reconstitution of quinone reduction and characterization of Escherichia coli fumarate reductase activity. J Biol Chem. 1986 Feb 5;261(4):1808–1814. [PubMed]
  • Chang AC, Cohen SN. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. [PMC free article] [PubMed]
  • Chippaux M, Bonnefoy-Orth V, Ratouchniak J, Pascal MC. Operon fusions in the nitrate reductase operon and study of the control gene nir R in Escherichia coli. Mol Gen Genet. 1981;182(3):477–479. [PubMed]
  • Chippaux M, Giudici D, Abou-Jaoudé A, Casse F, Pascal MC. Laboratoire de Chimie Bactérienne C.N.R.S., Marsielle, France. Mol Gen Genet. 1978 Apr 6;160(2):225–229. [PubMed]
  • Cole ST, Grundström T, Jaurin B, Robinson JJ, Weiner JH. Location and nucleotide sequence of frdB, the gene coding for the iron-sulphur protein subunit of the fumarate reductase of Escherichia coli. Eur J Biochem. 1982 Aug;126(1):211–216. [PubMed]
  • Cole ST, Guest JR. Genetic and physical characterization of lambda transducing phages (lambda frdA) containing the fumarate reductase gene of Escherichia coli K12. Mol Gen Genet. 1980;178(2):409–418. [PubMed]
  • Cowing DW, Bardwell JC, Craig EA, Woolford C, Hendrix RW, Gross CA. Consensus sequence for Escherichia coli heat shock gene promoters. Proc Natl Acad Sci U S A. 1985 May;82(9):2679–2683. [PMC free article] [PubMed]
  • de Crombrugghe B, Busby S, Buc H. Cyclic AMP receptor protein: role in transcription activation. Science. 1984 May 25;224(4651):831–838. [PubMed]
  • Dixon RA. The genetic complexity of nitrogen fixation. The ninth Fleming lecture. J Gen Microbiol. 1984 Nov;130(11):2745–2755. [PubMed]
  • Edlund T, Grundström T, Normark S. Isolation and characterization of DNA repetitions carrying the chromosomal beta-lactamase gene of Escherichia coli K-12. Mol Gen Genet. 1979 Jun 7;173(2):115–125. [PubMed]
  • Fimmel AL, Haddock BA. Use of chlC-lac fusions to determine regulation of gene chlC in Escherichia coli K-12. J Bacteriol. 1979 Jun;138(3):726–730. [PMC free article] [PubMed]
  • Fowler AV, Zabin I. Purification, structure, and properties of hybrid beta-galactosidase proteins. J Biol Chem. 1983 Dec 10;258(23):14354–14358. [PubMed]
  • Grundström T, Jaurin B. Overlap between ampC and frd operons on the Escherichia coli chromosome. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1111–1115. [PMC free article] [PubMed]
  • Guest JR. Partial replacement of succinate dehydrogenase function by phage- and plasmid-specified fumarate reductase in Escherichia coli. J Gen Microbiol. 1981 Feb;122(2):171–179. [PubMed]
  • HORIUCHI T, TOMIZAWA JI, NOVICK A. Isolation and properties of bacteria capable of high rates of beta-galactosidase synthesis. Biochim Biophys Acta. 1962 Jan 22;55:152–163. [PubMed]
  • Hunt TP, Magasanik B. Transcription of glnA by purified Escherichia coli components: core RNA polymerase and the products of glnF, glnG, and glnL. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8453–8457. [PMC free article] [PubMed]
  • Ingledew WJ, Poole RK. The respiratory chains of Escherichia coli. Microbiol Rev. 1984 Sep;48(3):222–271. [PMC free article] [PubMed]
  • Iuchi S, Kuritzkes DR, Lin EC. Escherichia coli mutant with altered respiratory control of the frd operon. J Bacteriol. 1985 Mar;161(3):1023–1028. [PMC free article] [PubMed]
  • Iuchi S, Kuritzkes DR, Lin EC. Three classes of Escherichia coli mutants selected for aerobic expression of fumarate reductase. J Bacteriol. 1986 Dec;168(3):1415–1421. [PMC free article] [PubMed]
  • Jamieson DJ, Sawers RG, Rugman PA, Boxer DH, Higgins CF. Effects of anaerobic regulatory mutations and catabolite repression on regulation of hydrogen metabolism and hydrogenase isoenzyme composition in Salmonella typhimurium. J Bacteriol. 1986 Oct;168(1):405–411. [PMC free article] [PubMed]
  • Jones HM, Gunsalus RP. Transcription of the Escherichia coli fumarate reductase genes (frdABCD) and their coordinate regulation by oxygen, nitrate, and fumarate. J Bacteriol. 1985 Dec;164(3):1100–1109. [PMC free article] [PubMed]
  • Kuritzkes DR, Zhang XY, Lin EC. Use of phi(glp-lac) in studies of respiratory regulation of the Escherichia coli anaerobic sn-glycerol-3-phosphate dehydrogenase genes (glpAB). J Bacteriol. 1984 Feb;157(2):591–598. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lambden PR, Guest JR. Mutants of Escherichia coli K12 unable to use fumarate as an anaerobic electron acceptor. J Gen Microbiol. 1976 Dec;97(2):145–160. [PubMed]
  • Lemire BD, Robinson JJ, Weiner JH. Identification of membrane anchor polypeptides of Escherichia coli fumarate reductase. J Bacteriol. 1982 Dec;152(3):1126–1131. [PMC free article] [PubMed]
  • Lohmeier E, Hagen DS, Dickie P, Weiner JH. Cloning and expression of fumarate reductase gene of Escherichia coli. Can J Biochem. 1981 Mar;59(3):158–164. [PubMed]
  • Merrick MJ, Gibbins JR. The nucleotide sequence of the nitrogen-regulation gene ntrA of Klebsiella pneumoniae and comparison with conserved features in bacterial RNA polymerase sigma factors. Nucleic Acids Res. 1985 Nov 11;13(21):7607–7620. [PMC free article] [PubMed]
  • Morningstar JE, Johnson MK, Cecchini G, Ackrell BA, Kearney EB. The high potential iron-sulfur center in Escherichia coli fumarate reductase is a three-iron cluster. J Biol Chem. 1985 Nov 5;260(25):13631–13638. [PubMed]
  • Newman BM, Cole JA. The chromosomal location and pleiotropic effects of mutations of the nirA+ gene of Escherichia coli K12: the essential role of nirA+ in nitrite reduction and in other anaerobic redox reactions. J Gen Microbiol. 1978 May;106(1):1–12. [PubMed]
  • Reddy P, Peterkofsky A, McKenney K. Translational efficiency of the Escherichia coli adenylate cyclase gene: mutating the UUG initiation codon to GUG or AUG results in increased gene expression. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5656–5660. [PMC free article] [PubMed]
  • Shaw DJ, Guest JR. Molecular cloning of the fnr gene of Escherichia coli K12. Mol Gen Genet. 1981;181(1):95–100. [PubMed]
  • Shaw DJ, Guest JR. Amplification and product identification of the fnr gene of Escherichia coli. J Gen Microbiol. 1982 Oct;128(10):2221–2228. [PubMed]
  • Shaw DJ, Guest JR. Nucleotide sequence of the fnr gene and primary structure of the Enr protein of Escherichia coli. Nucleic Acids Res. 1982 Oct 11;10(19):6119–6130. [PMC free article] [PubMed]
  • Shaw DJ, Rice DW, Guest JR. Homology between CAP and Fnr, a regulator of anaerobic respiration in Escherichia coli. J Mol Biol. 1983 May 15;166(2):241–247. [PubMed]
  • Simons RW, Houman F, Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. [PubMed]
  • Spencer ME, Guest JR. Isolation and properties of fumarate reductase mutants of Escherichia coli. J Bacteriol. 1973 May;114(2):563–570. [PMC free article] [PubMed]
  • Stragier P, Parsot C, Bouvier J. Two functional domains conserved in major and alternate bacterial sigma factors. FEBS Lett. 1985 Jul 22;187(1):11–15. [PubMed]
  • Stewart V. Requirement of Fnr and NarL functions for nitrate reductase expression in Escherichia coli K-12. J Bacteriol. 1982 Sep;151(3):1320–1325. [PMC free article] [PubMed]
  • Stewart V, MacGregor CH. Nitrate reductase in Escherichia coli K-12: involvement of chlC, chlE, and chlG loci. J Bacteriol. 1982 Aug;151(2):788–799. [PMC free article] [PubMed]
  • Strauch KL, Lenk JB, Gamble BL, Miller CG. Oxygen regulation in Salmonella typhimurium. J Bacteriol. 1985 Feb;161(2):673–680. [PMC free article] [PubMed]
  • Weiner JH, Dickie P. Fumarate reductase of Escherichia coli. Elucidation of the covalent-flavin component. J Biol Chem. 1979 Sep 10;254(17):8590–8593. [PubMed]

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