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. 1994 Apr;176(7):1831–1839. doi: 10.1128/jb.176.7.1831-1839.1994

Regulation of the Escherichia coli lrp gene.

Q Wang 1, J Wu 1, D Friedberg 1, J Plakto 1, J M Calvo 1
PMCID: PMC205284  PMID: 8144448

Abstract

Lrp (leucine-responsive regulatory protein) is a major Escherichia coli regulatory protein which regulates expression of a number of operons, some negatively and some positively. This work relates to a characterization of lrp, the gene encoding Lrp. Nucleotide sequencing established that the coding regions of lrp and trxB (encoding thioredoxin reductase) are separated by 543 bp and that the two genes are transcribed in opposite directions. In addition, we used primer extension, deletion analyses, and lrp-lacZ transcriptional fusions to delineate the promoter and regulatory region of the lrp operon. The lrp promoter is located 267 nucleotides upstream of the translational start codon of the lrp gene. In comparison with a wild-type strain, expression of the lrp operon was increased about 3-fold in a strain lacking Lrp and decreased about 10-fold in a strain overproducing Lrp. As observed from DNA mobility shift and DNase I footprinting analyses, Lrp binds to one or more sites within the region -80 to -32 relative to the start point of lrp transcription. A mutational analysis indicated that this same region is at least partly required for repression of lrp expression in vivo. These results demonstrate that autogenous regulation of lrp involves Lrp acting directly to cause repression of lrp transcription.

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Selected References

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  1. Braaten B. A., Platko J. V., van der Woude M. W., Simons B. H., de Graaf F. K., Calvo J. M., Low D. A. Leucine-responsive regulatory protein controls the expression of both the pap and fan pili operons in Escherichia coli. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4250–4254. doi: 10.1073/pnas.89.10.4250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ernsting B. R., Atkinson M. R., Ninfa A. J., Matthews R. G. Characterization of the regulon controlled by the leucine-responsive regulatory protein in Escherichia coli. J Bacteriol. 1992 Feb;174(4):1109–1118. doi: 10.1128/jb.174.4.1109-1118.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Gazeau M., Delort F., Dessen P., Blanquet S., Plateau P. Escherichia coli leucine-responsive regulatory protein (Lrp) controls lysyl-tRNA synthetase expression. FEBS Lett. 1992 Apr 6;300(3):254–258. doi: 10.1016/0014-5793(92)80857-d. [DOI] [PubMed] [Google Scholar]
  4. Haney S. A., Platko J. V., Oxender D. L., Calvo J. M. Lrp, a leucine-responsive protein, regulates branched-chain amino acid transport genes in Escherichia coli. J Bacteriol. 1992 Jan;174(1):108–115. doi: 10.1128/jb.174.1.108-115.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kölling R., Lother H. AsnC: an autogenously regulated activator of asparagine synthetase A transcription in Escherichia coli. J Bacteriol. 1985 Oct;164(1):310–315. doi: 10.1128/jb.164.1.310-315.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Lewis L. K., Jenkins M. E., Mount D. W. Isolation of DNA damage-inducible promoters in Escherichia coli: regulation of polB (dinA), dinG, and dinH by LexA repressor. J Bacteriol. 1992 May;174(10):3377–3385. doi: 10.1128/jb.174.10.3377-3385.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lin R., D'Ari R., Newman E. B. Lambda placMu insertions in genes of the leucine regulon: extension of the regulon to genes not regulated by leucine. J Bacteriol. 1992 Mar;174(6):1948–1955. doi: 10.1128/jb.174.6.1948-1955.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lin R., Ernsting B., Hirshfield I. N., Matthews R. G., Neidhardt F. C., Clark R. L., Newman E. B. The lrp gene product regulates expression of lysU in Escherichia coli K-12. J Bacteriol. 1992 May;174(9):2779–2784. doi: 10.1128/jb.174.9.2779-2784.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Mulligan M. E., Hawley D. K., Entriken R., McClure W. R. Escherichia coli promoter sequences predict in vitro RNA polymerase selectivity. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):789–800. doi: 10.1093/nar/12.1part2.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Newman E. B., D'Ari R., Lin R. T. The leucine-Lrp regulon in E. coli: a global response in search of a raison d'être. Cell. 1992 Feb 21;68(4):617–619. doi: 10.1016/0092-8674(92)90135-y. [DOI] [PubMed] [Google Scholar]
  11. Nou X., Skinner B., Braaten B., Blyn L., Hirsch D., Low D. Regulation of pyelonephritis-associated pili phase-variation in Escherichia coli: binding of the PapI and the Lrp regulatory proteins is controlled by DNA methylation. Mol Microbiol. 1993 Feb;7(4):545–553. doi: 10.1111/j.1365-2958.1993.tb01145.x. [DOI] [PubMed] [Google Scholar]
  12. Platko J. V., Calvo J. M. Mutations affecting the ability of Escherichia coli Lrp to bind DNA, activate transcription, or respond to leucine. J Bacteriol. 1993 Feb;175(4):1110–1117. doi: 10.1128/jb.175.4.1110-1117.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Platko J. V., Willins D. A., Calvo J. M. The ilvIH operon of Escherichia coli is positively regulated. J Bacteriol. 1990 Aug;172(8):4563–4570. doi: 10.1128/jb.172.8.4563-4570.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rex J. H., Aronson B. D., Somerville R. L. The tdh and serA operons of Escherichia coli: mutational analysis of the regulatory elements of leucine-responsive genes. J Bacteriol. 1991 Oct;173(19):5944–5953. doi: 10.1128/jb.173.19.5944-5953.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rosenthal E. R., Calvo J. M. Transcription termination sites at the distal end of the leu operon of Salmonella typhimurium. J Mol Biol. 1987 Apr 5;194(3):443–452. doi: 10.1016/0022-2836(87)90673-5. [DOI] [PubMed] [Google Scholar]
  16. Ross W., Gosink K. K., Salomon J., Igarashi K., Zou C., Ishihama A., Severinov K., Gourse R. L. A third recognition element in bacterial promoters: DNA binding by the alpha subunit of RNA polymerase. Science. 1993 Nov 26;262(5138):1407–1413. doi: 10.1126/science.8248780. [DOI] [PubMed] [Google Scholar]
  17. Russel M., Model P. Sequence of thioredoxin reductase from Escherichia coli. Relationship to other flavoprotein disulfide oxidoreductases. J Biol Chem. 1988 Jun 25;263(18):9015–9019. [PubMed] [Google Scholar]
  18. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Tuan L. R., D'Ari R., Newman E. B. The leucine regulon of Escherichia coli K-12: a mutation in rblA alters expression of L-leucine-dependent metabolic operons. J Bacteriol. 1990 Aug;172(8):4529–4535. doi: 10.1128/jb.172.8.4529-4535.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wang Q., Calvo J. M. Lrp, a global regulatory protein of Escherichia coli, binds co-operatively to multiple sites and activates transcription of ilvIH. J Mol Biol. 1993 Jan 20;229(2):306–318. doi: 10.1006/jmbi.1993.1036. [DOI] [PubMed] [Google Scholar]
  22. Wang Q., Calvo J. M. Lrp, a major regulatory protein in Escherichia coli, bends DNA and can organize the assembly of a higher-order nucleoprotein structure. EMBO J. 1993 Jun;12(6):2495–2501. doi: 10.1002/j.1460-2075.1993.tb05904.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wang Q., Sacco M., Ricca E., Lago C. T., De Felice M., Calvo J. M. Organization of Lrp-binding sites upstream of ilvIH in Salmonella typhimurium. Mol Microbiol. 1993 Mar;7(6):883–891. doi: 10.1111/j.1365-2958.1993.tb01179.x. [DOI] [PubMed] [Google Scholar]
  24. Willins D. A., Calvo J. M. In vitro transcription from the Escherichia coli ilvIH promoter. J Bacteriol. 1992 Dec;174(23):7648–7655. doi: 10.1128/jb.174.23.7648-7655.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Willins D. A., Ryan C. W., Platko J. V., Calvo J. M. Characterization of Lrp, and Escherichia coli regulatory protein that mediates a global response to leucine. J Biol Chem. 1991 Jun 15;266(17):10768–10774. [PubMed] [Google Scholar]
  26. de Wind N., de Jong M., Meijer M., Stuitje A. R. Site-directed mutagenesis of the Escherichia coli chromosome near oriC: identification and characterization of asnC, a regulatory element in E. coli asparagine metabolism. Nucleic Acids Res. 1985 Dec 20;13(24):8797–8811. doi: 10.1093/nar/13.24.8797. [DOI] [PMC free article] [PubMed] [Google Scholar]

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