Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1994 Sep;176(17):5270–5276. doi: 10.1128/jb.176.17.5270-5276.1994

Effect of heme and oxygen availability on hemA gene expression in Escherichia coli: role of the fnr, arcA, and himA gene products.

S Darie 1, R P Gunsalus 1
PMCID: PMC196710  PMID: 8071201

Abstract

While many organisms synthesize delta-aminolevulinate, the precursor of heme, by condensing succinyl-coenzyme A and glycine, others use a glutamate-dependent pathway in which glutamyl-tRNA dehydrogenase catalyzes the rate-determining step. The hemeA gene that encodes this latter enzyme in Escherichia coli has been cloned and sequenced. To examine how its expression is regulated, we constructed hemA-lacZ operon and gene fusions and inserted them into the chromosome in single copy. The effect of aerobic and anaerobic growth conditions and the availability of electron acceptors and various carbon substrates were documented. Use of different types of cell culture medium resulted in a fivefold variation in hemA-lacZ expression during aerobic cell growth. Anaerobic growth resulted in 2.5-fold-higher hemA-lacZ expression than aerobic growth. This control is mediated by the fnr and arcA gene products. Fnr functions as a repressor of hemA transcription during anaerobic cell growth only, whereas the arcA gene product activates hemA gene expression under both aerobic and anaerobic conditions. Integration host factor protein was also shown to be required for control of hemA gene regulation. To determine whether an intermediate or a product of the heme biosynthetic pathway is involved in hemA regulation, hemA-lacZ expression was analyzed in a hemA mutant. Expression was elevated by 20-fold compared with that in a wild-type strain, while the addition of the heme pathway intermediate delta-aminolevulinate to the culture medium restored expression to wild-type levels. These results suggest that the heme pathway is feedback regulated at the level of hemA gene expression, to supply heme as it is required during different modes of cell growth.

Full text

PDF
5274

Selected References

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

  1. Avissar Y. J., Beale S. I. Identification of the enzymatic basis for delta-aminolevulinic acid auxotrophy in a hemA mutant of Escherichia coli. J Bacteriol. 1989 Jun;171(6):2919–2924. doi: 10.1128/jb.171.6.2919-2924.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cotter P. A., Chepuri V., Gennis R. B., Gunsalus R. P. Cytochrome o (cyoABCDE) and d (cydAB) oxidase gene expression in Escherichia coli is regulated by oxygen, pH, and the fnr gene product. J Bacteriol. 1990 Nov;172(11):6333–6338. doi: 10.1128/jb.172.11.6333-6338.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cotter P. A., Darie S., Gunsalus R. P. The effect of iron limitation on expression of the aerobic and anaerobic electron transport pathway genes in Escherichia coli. FEMS Microbiol Lett. 1992 Dec 15;100(1-3):227–232. doi: 10.1111/j.1574-6968.1992.tb14045.x. [DOI] [PubMed] [Google Scholar]
  4. Cotter P. A., Gunsalus R. P. Contribution of the fnr and arcA gene products in coordinate regulation of cytochrome o and d oxidase (cyoABCDE and cydAB) genes in Escherichia coli. FEMS Microbiol Lett. 1992 Feb 1;70(1):31–36. doi: 10.1016/0378-1097(92)90558-6. [DOI] [PubMed] [Google Scholar]
  5. Cotter P. A., Gunsalus R. P. Oxygen, nitrate, and molybdenum regulation of dmsABC gene expression in Escherichia coli. J Bacteriol. 1989 Jul;171(7):3817–3823. doi: 10.1128/jb.171.7.3817-3823.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. De Lorenzo V., Herrero M., Giovannini F., Neilands J. B. Fur (ferric uptake regulation) protein and CAP (catabolite-activator protein) modulate transcription of fur gene in Escherichia coli. Eur J Biochem. 1988 May 2;173(3):537–546. doi: 10.1111/j.1432-1033.1988.tb14032.x. [DOI] [PubMed] [Google Scholar]
  7. Drolet M., Péloquin L., Echelard Y., Cousineau L., Sasarman A. Isolation and nucleotide sequence of the hemA gene of Escherichia coli K12. Mol Gen Genet. 1989 Apr;216(2-3):347–352. doi: 10.1007/BF00334375. [DOI] [PubMed] [Google Scholar]
  8. Friedman D. I. Integration host factor: a protein for all reasons. Cell. 1988 Nov 18;55(4):545–554. doi: 10.1016/0092-8674(88)90213-9. [DOI] [PubMed] [Google Scholar]
  9. Fu H. A., Iuchi S., Lin E. C. The requirement of ArcA and Fnr for peak expression of the cyd operon in Escherichia coli under microaerobic conditions. Mol Gen Genet. 1991 Apr;226(1-2):209–213. doi: 10.1007/BF00273605. [DOI] [PubMed] [Google Scholar]
  10. Grove C. L., Gunsalus R. P. Regulation of the aroH operon of Escherichia coli by the tryptophan repressor. J Bacteriol. 1987 May;169(5):2158–2164. doi: 10.1128/jb.169.5.2158-2164.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gunsalus R. P. Control of electron flow in Escherichia coli: coordinated transcription of respiratory pathway genes. J Bacteriol. 1992 Nov;174(22):7069–7074. doi: 10.1128/jb.174.22.7069-7074.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gunsalus R. P., Zurawski G., Yanofsky C. Structural and functional analysis of cloned deoxyribonucleic acid containing the trpR-thr region of the Escherichia coli chromosome. J Bacteriol. 1979 Oct;140(1):106–113. doi: 10.1128/jb.140.1.106-113.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Iuchi S., Lin E. C. Adaptation of Escherichia coli to respiratory conditions: regulation of gene expression. Cell. 1991 Jul 12;66(1):5–7. doi: 10.1016/0092-8674(91)90130-q. [DOI] [PubMed] [Google Scholar]
  14. Jahn D., Michelsen U., Söll D. Two glutamyl-tRNA reductase activities in Escherichia coli. J Biol Chem. 1991 Feb 5;266(4):2542–2548. [PubMed] [Google Scholar]
  15. Johnson R. C., Ball C. A., Pfeffer D., Simon M. I. Isolation of the gene encoding the Hin recombinational enhancer binding protein. Proc Natl Acad Sci U S A. 1988 May;85(10):3484–3488. doi: 10.1073/pnas.85.10.3484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Li J. M., Brathwaite O., Cosloy S. D., Russell C. S. 5-Aminolevulinic acid synthesis in Escherichia coli. J Bacteriol. 1989 May;171(5):2547–2552. doi: 10.1128/jb.171.5.2547-2552.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Li J. M., Russell C. S., Cosloy S. D. Cloning and structure of the hem A gene of Escherichia coli K-12. Gene. 1989 Oct 30;82(2):209–217. doi: 10.1016/0378-1119(89)90046-2. [DOI] [PubMed] [Google Scholar]
  18. Niehaus F., Hantke K., Unden G. Iron content and FNR-dependent gene regulation in Escherichia coli. FEMS Microbiol Lett. 1991 Dec 1;68(3):319–323. doi: 10.1016/0378-1097(91)90376-l. [DOI] [PubMed] [Google Scholar]
  19. Padmanaban G., Venkateswar V., Rangarajan P. N. Haem as a multifunctional regulator. Trends Biochem Sci. 1989 Dec;14(12):492–496. doi: 10.1016/0968-0004(89)90182-5. [DOI] [PubMed] [Google Scholar]
  20. Petricek M., Rutberg L., Schröder I., Hederstedt L. Cloning and characterization of the hemA region of the Bacillus subtilis chromosome. J Bacteriol. 1990 May;172(5):2250–2258. doi: 10.1128/jb.172.5.2250-2258.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rice C. W., Hempfling W. P. Oxygen-limited continuous culture and respiratory energy conservation in Escherichia coli. J Bacteriol. 1978 Apr;134(1):115–124. doi: 10.1128/jb.134.1.115-124.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rimington C. Haem biosynthesis and porphyrias: 50 years in retrospect. J Clin Chem Clin Biochem. 1989 Aug;27(8):473–486. [PubMed] [Google Scholar]
  23. Schröder I., Darie S., Gunsalus R. P. Activation of the Escherichia coli nitrate reductase (narGHJI) operon by NarL and Fnr requires integration host factor. J Biol Chem. 1993 Jan 15;268(2):771–774. [PubMed] [Google Scholar]
  24. Simons R. W., Houman F., Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. doi: 10.1016/0378-1119(87)90095-3. [DOI] [PubMed] [Google Scholar]
  25. Spiro S., Guest J. R. FNR and its role in oxygen-regulated gene expression in Escherichia coli. FEMS Microbiol Rev. 1990 Aug;6(4):399–428. doi: 10.1111/j.1574-6968.1990.tb04109.x. [DOI] [PubMed] [Google Scholar]
  26. Săsărman A., Surdeanu M., Horodniceanu T. Locus determining the synthesis of delta-aminolevulinic acid in Escherichia coli K-12. J Bacteriol. 1968 Nov;96(5):1882–1884. doi: 10.1128/jb.96.5.1882-1884.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Verkamp E., Backman V. M., Björnsson J. M., Söll D., Eggertsson G. The periplasmic dipeptide permease system transports 5-aminolevulinic acid in Escherichia coli. J Bacteriol. 1993 Mar;175(5):1452–1456. doi: 10.1128/jb.175.5.1452-1456.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Verkamp E., Chelm B. K. Isolation, nucleotide sequence, and preliminary characterization of the Escherichia coli K-12 hemA gene. J Bacteriol. 1989 Sep;171(9):4728–4735. doi: 10.1128/jb.171.9.4728-4735.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Verkamp E., Jahn M., Jahn D., Kumar A. M., Söll D. Glutamyl-tRNA reductase from Escherichia coli and Synechocystis 6803. Gene structure and expression. J Biol Chem. 1992 Apr 25;267(12):8275–8280. [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES