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. 1996 Feb;178(3):638–646. doi: 10.1128/jb.178.3.638-646.1996

Transcription of the glutamyl-tRNA reductase (hemA) gene in Salmonella typhimurium and Escherichia coli: role of the hemA P1 promoter and the arcA gene product.

P Choi 1, L Wang 1, C D Archer 1, T Elliott 1
PMCID: PMC177706  PMID: 8550494

Abstract

In Salmonella typhimurium and Escherichia coli, the hemA gene encodes the enzyme glutamyl-tRNA reductase, which catalyzes the first committed step in the heme biosynthetic pathway. It has recently been reported that a lac operon fusion to the hemA promoter of E. coli is induced 20-fold after starvation for heme. Induction was dependent on the transcriptional regulator ArcA, with a second transcriptional regulator, FNR, playing a negative role specifically under anaerobic conditions (S. Darie and R. P. Gunsalus, J. Bacteriol. 176:5270-5276, 1994). We have investigated the generality of this effect by examining the response to heme starvation of a number of lac operon fusions to the hemA promoters of both E. coli and S. typhimurium. We confirmed that such fusions are induced during starvation of a hemA auxotroph, but the level of induction observed was maximally sixfold and for S. typhimurium fusions it was only two- to fourfold. Sequences required for high-level expression of hemA lie within 129 bp upstream of the major (P1) promoter transcriptional start site. Mutants defective in the P1 promoter had greatly reduced hemA-lac expression both in the presence and in the absence of ALA. Mutations in arcA had no effect on hemA-lac expression in E. coli during normal growth, although the increase in expression during starvation for ALA was half that seen in an arcA+ strain. Overexpression of the arcA gene had no effect on hemA-lac expression. Primer extension analysis showed that RNA 5' ends mapping to the hemA P1 and P2 promoters were not expressed at significantly higher levels in induced cultures. These results differ from those previously reported.

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Selected References

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  1. Ailion M., Bobik T. A., Roth J. R. Two global regulatory systems (Crp and Arc) control the cobalamin/propanediol regulon of Salmonella typhimurium. J Bacteriol. 1993 Nov;175(22):7200–7208. doi: 10.1128/jb.175.22.7200-7208.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Avissar Y. J., Beale S. I. Cloning and expression of a structural gene from Chlorobium vibrioforme that complements the hemA mutation in Escherichia coli. J Bacteriol. 1990 Mar;172(3):1656–1659. doi: 10.1128/jb.172.3.1656-1659.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Avissar Y. J., Ormerod J. G., Beale S. I. Distribution of delta-aminolevulinic acid biosynthetic pathways among phototrophic bacterial groups. Arch Microbiol. 1989;151(6):513–519. doi: 10.1007/BF00454867. [DOI] [PubMed] [Google Scholar]
  5. Chen W., Russell C. S., Murooka Y., Cosloy S. D. 5-Aminolevulinic acid synthesis in Escherichia coli requires expression of hemA. J Bacteriol. 1994 May;176(9):2743–2746. doi: 10.1128/jb.176.9.2743-2746.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Darie S., Gunsalus R. P. Effect of heme and oxygen availability on hemA gene expression in Escherichia coli: role of the fnr, arcA, and himA gene products. J Bacteriol. 1994 Sep;176(17):5270–5276. doi: 10.1128/jb.176.17.5270-5276.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Elliott T. A method for constructing single-copy lac fusions in Salmonella typhimurium and its application to the hemA-prfA operon. J Bacteriol. 1992 Jan;174(1):245–253. doi: 10.1128/jb.174.1.245-253.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Elliott T., Avissar Y. J., Rhie G. E., Beale S. I. Cloning and sequence of the Salmonella typhimurium hemL gene and identification of the missing enzyme in hemL mutants as glutamate-1-semialdehyde aminotransferase. J Bacteriol. 1990 Dec;172(12):7071–7084. doi: 10.1128/jb.172.12.7071-7084.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Elliott T. Cloning, genetic characterization, and nucleotide sequence of the hemA-prfA operon of Salmonella typhimurium. J Bacteriol. 1989 Jul;171(7):3948–3960. doi: 10.1128/jb.171.7.3948-3960.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Elliott T., Roth J. R. Heme-deficient mutants of Salmonella typhimurium: two genes required for ALA synthesis. Mol Gen Genet. 1989 Apr;216(2-3):303–314. doi: 10.1007/BF00334369. [DOI] [PubMed] [Google Scholar]
  11. Hart R. A., Kallio P. T., Bailey J. E. Effect of biosynthetic manipulation of heme on insolubility of Vitreoscilla hemoglobin in Escherichia coli. Appl Environ Microbiol. 1994 Jul;60(7):2431–2437. doi: 10.1128/aem.60.7.2431-2437.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Herlitze S., Koenen M. A general and rapid mutagenesis method using polymerase chain reaction. Gene. 1990 Jul 2;91(1):143–147. doi: 10.1016/0378-1119(90)90177-s. [DOI] [PubMed] [Google Scholar]
  13. Hino S., Ishida A. Effect of oxygen on heme and cytochrome content in some facultative bacteria. Enzyme. 1973;16(1):42–49. doi: 10.1159/000459360. [DOI] [PubMed] [Google Scholar]
  14. Iuchi S., Lin E. C. arcA (dye), a global regulatory gene in Escherichia coli mediating repression of enzymes in aerobic pathways. Proc Natl Acad Sci U S A. 1988 Mar;85(6):1888–1892. doi: 10.1073/pnas.85.6.1888. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Jahn D., Verkamp E., Söll D. Glutamyl-transfer RNA: a precursor of heme and chlorophyll biosynthesis. Trends Biochem Sci. 1992 Jun;17(6):215–218. doi: 10.1016/0968-0004(92)90380-r. [DOI] [PubMed] [Google Scholar]
  17. Janzer J. J., Stan-Lotter H., Sanderson K. E. Isolation and characterization of hemin-permeable, envelope-defective mutants of Salmonella typhimurium. Can J Microbiol. 1981 Feb;27(2):226–237. doi: 10.1139/m81-034. [DOI] [PubMed] [Google Scholar]
  18. Lazazzera B. A., Bates D. M., Kiley P. J. The activity of the Escherichia coli transcription factor FNR is regulated by a change in oligomeric state. Genes Dev. 1993 Oct;7(10):1993–2005. doi: 10.1101/gad.7.10.1993. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Mogi T., Saiki K., Anraku Y. Biosynthesis and functional role of haem O and haem A. Mol Microbiol. 1994 Nov;14(3):391–398. doi: 10.1111/j.1365-2958.1994.tb02174.x. [DOI] [PubMed] [Google Scholar]
  21. Morrison T. B., Parkinson J. S. Liberation of an interaction domain from the phosphotransfer region of CheA, a signaling kinase of Escherichia coli. Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5485–5489. doi: 10.1073/pnas.91.12.5485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. O'Neill G. P., Chen M. W., Söll D. delta-Aminolevulinic acid biosynthesis in Escherichia coli and Bacillus subtilis involves formation of glutamyl-tRNA. FEMS Microbiol Lett. 1989 Aug;51(3):255–259. doi: 10.1016/0378-1097(89)90406-0. [DOI] [PubMed] [Google Scholar]
  23. Post D. A., Hove-Jensen B., Switzer R. L. Characterization of the hemA-prs region of the Escherichia coli and Salmonella typhimurium chromosomes: identification of two open reading frames and implications for prs expression. J Gen Microbiol. 1993 Feb;139(2):259–266. doi: 10.1099/00221287-139-2-259. [DOI] [PubMed] [Google Scholar]
  24. RICHMOND M. H., MALLOE O. The rate of growth of Salmonella typhimurium with individual carbon sources related to glucose metabolism or to the Krebs cycle. J Gen Microbiol. 1962 Feb;27:285–297. doi: 10.1099/00221287-27-2-285. [DOI] [PubMed] [Google Scholar]
  25. Roeder W., Somerville R. L. Cloning the trpR gene. Mol Gen Genet. 1979 Nov;176(3):361–368. doi: 10.1007/BF00333098. [DOI] [PubMed] [Google Scholar]
  26. Schmieger H. Phage P22-mutants with increased or decreased transduction abilities. Mol Gen Genet. 1972;119(1):75–88. doi: 10.1007/BF00270447. [DOI] [PubMed] [Google Scholar]
  27. 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]
  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]
  30. Warren M. J., Bolt E. L., Roessner C. A., Scott A. I., Spencer J. B., Woodcock S. C. Gene dissection demonstrates that the Escherichia coli cysG gene encodes a multifunctional protein. Biochem J. 1994 Sep 15;302(Pt 3):837–844. doi: 10.1042/bj3020837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wilmes-Riesenberg M. R., Wanner B. L. TnphoA and TnphoA' elements for making and switching fusions for study of transcription, translation, and cell surface localization. J Bacteriol. 1992 Jul;174(14):4558–4575. doi: 10.1128/jb.174.14.4558-4575.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wilson H. R., Archer C. D., Liu J. K., Turnbough C. L., Jr Translational control of pyrC expression mediated by nucleotide-sensitive selection of transcriptional start sites in Escherichia coli. J Bacteriol. 1992 Jan;174(2):514–524. doi: 10.1128/jb.174.2.514-524.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Xu K., Delling J., Elliott T. The genes required for heme synthesis in Salmonella typhimurium include those encoding alternative functions for aerobic and anaerobic coproporphyrinogen oxidation. J Bacteriol. 1992 Jun;174(12):3953–3963. doi: 10.1128/jb.174.12.3953-3963.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Xu K., Elliott T. An oxygen-dependent coproporphyrinogen oxidase encoded by the hemF gene of Salmonella typhimurium. J Bacteriol. 1993 Aug;175(16):4990–4999. doi: 10.1128/jb.175.16.4990-4999.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Xu K., Elliott T. Cloning, DNA sequence, and complementation analysis of the Salmonella typhimurium hemN gene encoding a putative oxygen-independent coproporphyrinogen III oxidase. J Bacteriol. 1994 Jun;176(11):3196–3203. doi: 10.1128/jb.176.11.3196-3203.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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