1887

Abstract

In this study, the gene encoding the leucine-responsive regulatory protein (Lrp) in serovar Typhimurium was found to be negatively autoregulated. Its transcription start site was determined by primer extension analysis, showing that the promoter is located at a different site to that inferred previously from the Typhimurium genome sequence. Chromosomal DNA fragments that include the promoter region were bound by purified Lrp protein , producing up to four distinct protein–DNA complexes. DNase I footprinting identified regions that were protected by the protein as well as bases that became hypersensitive to DNase I treatment following Lrp binding. A clear pattern of periodic hypersensitivity was detected between positions −130 and +15 that was consistent with wrapping of the DNA around Lrp in a nucleoprotein complex that includes the putative promoter region. Lrp–DNA interaction in this region was fully consistent with the observed repression of transcription by this protein. Leucine was found to modulate Lrp-mediated autorepression by remodelling the Lrp–DNA nucleoprotein complex.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.2008/018358-0
2008-07-01
2019-11-21
Loading full text...

Full text loading...

/deliver/fulltext/micro/154/7/2008.html?itemId=/content/journal/micro/10.1099/mic.0.2008/018358-0&mimeType=html&fmt=ahah

References

  1. Bell, S. D. & Jackson, S. P. ( 2000; ). Mechanism of autoregulation by an archaeal transcriptional repressor. J Biol Chem 275, 31624–31629.[CrossRef]
    [Google Scholar]
  2. Borst, D. W., Blumenthal, R. M. & Matthews, R. G. ( 1996; ). Use of an in vivo titration method to study a global regulator: effect of varying Lrp levels on expression of gltBDF in Escherichia coli. J Bacteriol 178, 6904–6912.
    [Google Scholar]
  3. Bouloc, P., Vinella, D. & D'Ari, R. ( 1992; ). Leucine and serine induce mecillinam resistance in Escherichia coli. Mol Gen Genet 235, 242–246.[CrossRef]
    [Google Scholar]
  4. Braaten, B. A., Platko, J. V., van der Woude, M. W., Simons, B. H., de Graaf, F. K., Calvo, J. M. & Low, D. A. ( 1992; ). Leucine-responsive regulatory protein controls the expression of both pap and fan pili operons in Escherichia coli. Proc Natl Acad Sci U S A 89, 4250–4254.[CrossRef]
    [Google Scholar]
  5. Brinkman, A. B., Bell, S. D., Lebbink, R. J., de Vos, W. M. & van der Oost, J. ( 2002; ). The Sulfolobus solfataricus Lrp-like protein LysM regulates lysine biosynthesis in response to lysine availability. J Biol Chem 277, 29537–29549.[CrossRef]
    [Google Scholar]
  6. Brinkman, A. B., Ettema, T. J. G., de Vos, W. M. & van der Oost, J. ( 2003; ). The Lrp family of transcriptional regulators. Mol Microbiol 48, 287–294.[CrossRef]
    [Google Scholar]
  7. Calvo, J. M. & Matthews, R. G. ( 1994; ). The leucine-responsive regulatory protein, a global regulator of metabolism in Escherichia coli. Microbiol Rev 58, 466–490.
    [Google Scholar]
  8. Camacho, E. M. & Casadesús, J. ( 2002; ). Conjugal transfer of the virulence plasmid of Salmonella enterica is regulated by the leucine-responsive regulatory protein and DNA adenine methylation. Mol Microbiol 44, 1589–1598.[CrossRef]
    [Google Scholar]
  9. Camacho, E. M. & Casadesús, J. ( 2005; ). Regulation of traJ transcription in the Salmonella virulence plasmid by strand-specific DNA adenine hemimethylation. Mol Microbiol 57, 1700–1718.[CrossRef]
    [Google Scholar]
  10. Chen, S. & Calvo, J. M. ( 2002; ). Leucine-induced dissociation of Escherichia coli Lrp hexadecamers to octamers. J Mol Biol 318, 1031–1042.[CrossRef]
    [Google Scholar]
  11. Chen, S., Rosner, M. H. & Calvo, J. M. ( 2001; ). Leucine-regulated self-association of leucine-responsive regulatory protein (Lrp) from Escherichia coli. J Mol Biol 312, 625–635.[CrossRef]
    [Google Scholar]
  12. Chen, S., Iannolo, M. & Calvo, J. M. ( 2005; ). Cooperative binding of the leucine-responsive regulatory protein (Lrp) to DNA. J Mol Biol 345, 251–264.[CrossRef]
    [Google Scholar]
  13. de los Rios, S. & Perona, J. J. ( 2007; ). Structure of the Escherichia coli leucine-responsive regulatory protein (Lrp) reveals a novel octameric assembly. J Mol Biol 366, 1589–1602.[CrossRef]
    [Google Scholar]
  14. Hautefort, I., Proenca, M. J. & Hinton, J. C. D. ( 2003; ). Single-copy green fluorescent protein gene fusions allow accurate measurement of Salmonella gene expression in vitro and during infection of mammalian cells. Appl Environ Microbiol 69, 7480–7491.[CrossRef]
    [Google Scholar]
  15. Hernday, A., Krabbe, M., Braaten, B. & Low, D. ( 2002; ). Self-perpetuating epigenetic pili switches in bacteria. Proc Natl Acad Sci U S A 99, 16470–16476.[CrossRef]
    [Google Scholar]
  16. Hoiseth, S. K. & Stocker, B. A. ( 1981; ). Aromatic-dependent Salmonella typhimurium are non-virulent and effective as live vaccines. Nature 291, 238–239.[CrossRef]
    [Google Scholar]
  17. Kelly, A., Conway, C., Cróinín, T. O., Smith, S. G. J. & Dorman, C. J. ( 2006; ). DNA supercoiling and the Lrp protein determine the directionality of fim switch DNA inversion in Escherichia coli K-12. J Bacteriol 188, 5356–5363.[CrossRef]
    [Google Scholar]
  18. Landgraf, J. R., Wu, J. & Calvo, J. ( 1996; ). Effects of nutrition and growth rate on Lrp levels in Escherichia coli. J Bacteriol 178, 6930–6936.
    [Google Scholar]
  19. Lerner, C. G. & Inouye, M. ( 1990; ). Low copy number plasmids for regulated low-level expression of cloned genes in Escherichia coli with blue/white insert screening capability. Nucleic Acids Res 18, 4631 [CrossRef]
    [Google Scholar]
  20. Libby, S. J., Lesnick, M., Hasegawa, P., Weidenhammer, E. & Guiney, D. G. ( 2000; ). The Salmonella virulence plasmid spv genes are required for cytopathology in human monocyte-derived macrophages. Cell Microbiol 2, 49–58.[CrossRef]
    [Google Scholar]
  21. Lin, R., D'Ari, R. & Newman, E. B. ( 1992; ). λ placMu insertions in genes of the leucine regulon: extension of the regulon to genes not regulated by leucine. J Bacteriol 174, 1948–1955.
    [Google Scholar]
  22. Marshall, D. G., Sheehan, B. J. & Dorman, C. J. ( 1999; ). A role for the leucine-responsive regulatory protein and integration host factor in the regulation of the Salmonella plasmid virulence (spv) locus in Salmonella typhimurium. Mol Microbiol 34, 134–145.[CrossRef]
    [Google Scholar]
  23. McClelland, M., Sanderson, K. E., Spieth, J., Clifton, S. W., Latreille, P., Courtney, L., Porwollik, S., Ali, J., Dante, M. & other authors ( 2001; ). Complete genome sequence of Salmonella enterica serovar Typhimurium LT2. Nature 413, 852–856.[CrossRef]
    [Google Scholar]
  24. McCusker, M. P., Turner, E. C. & Dorman, C. J. ( 2008; ). DNA sequence heterogeneity in Fim tyrosine-integrase recombinase-binding elements and functional motif asymmetries determine the directionality of the fim genetic switch in Escherichia coli K-12. Mol Microbiol 67, 171–187.
    [Google Scholar]
  25. McFarland, K. A., Lucchini, S., Hinton, J. C. D. & Dorman, C. J. ( 2008; ). The leucine-responsive regulatory protein, Lrp, activates transcription of the fim operon in Salmonella enterica serovar Typhimurium via the fimZ regulatory gene. J Bacteriol 190, 602–612.[CrossRef]
    [Google Scholar]
  26. Nicholson, B. & Low, D. ( 2000; ). DNA methylation-dependent regulation of pef expression in Salmonella typhimurium. Mol Microbiol 35, 728–742.[CrossRef]
    [Google Scholar]
  27. Nickerson, C. A. & Achberger, E. C. ( 1995; ). Role of curved DNA in binding of Escherichia coli RNA polymerase to promoters. J Bacteriol 177, 5756–5761.
    [Google Scholar]
  28. Paesold, G., Guiney, D. G., Eckmann, L. & Kagnoff, M. F. ( 2002; ). Genes in the Salmonella pathogenicity island 2 and the Salmonella virulence plasmid are essential for Salmonella-induced apoptosis in intestinal epithelial cells. Cell Microbiol 4, 771–781.[CrossRef]
    [Google Scholar]
  29. Peterson, S. N., Dahlquist, F. W. & Reich, N. O. ( 2007; ). The role of high affinity non-specific DNA binding by Lrp in transcriptional regulation and DNA organization. J Mol Biol 369, 1307–1317.[CrossRef]
    [Google Scholar]
  30. Roesch, P. L. & Blomfield, I. C. ( 1998; ). Leucine alters the interaction of the leucine-responsive regulatory protein (Lrp) with the fim switch to stimulate site-specific recombination in Escherichia coli. Mol Microbiol 27, 751–761.[CrossRef]
    [Google Scholar]
  31. Rotger, R. & Casadesús, J. ( 1999; ). The virulence plasmids of Salmonella. Int Microbiol 2, 177–184.
    [Google Scholar]
  32. Tuan, L. R., D'Ari, R. & Newman, E. B. ( 1990; ). The leucine regulon of Escherichia coli K-12: a mutation in rblA alters expression of l-leucine-dependent metabolic operons. J Bacteriol 172, 4529–4535.
    [Google Scholar]
  33. van der Woude, M. W., Braaten, B. A. & Low, D. A. ( 1992; ). Evidence for global regulatory control of pilus expression in Escherichia coli by Lrp and DNA methylation: model building based on analysis of pap. Mol Microbiol 6, 2429–2435.
    [Google Scholar]
  34. Wagner, R. ( 2000; ). Transcription Regulation in Prokaryotes. Oxford, UK: Oxford University Press.
  35. Wang, Q. & Calvo, J. M. ( 1993; ). Lrp, a major regulatory protein in Escherichia coli, bends DNA and can organize the assembly of a higher-order nucleoprotein structure. EMBO J 12, 2495–2501.
    [Google Scholar]
  36. Wang, Q., Wu, J., Friedberg, D., Platko, J. & Calvo, J. M. ( 1994; ). Regulation of the Escherichia coli lrp gene. J Bacteriol 176, 1831–1839.
    [Google Scholar]
  37. Wiese, D. E., Jr, Ernsting, B. R., Blumenthal, R. M. & Matthews, R. G. ( 1997; ). A nucleoprotein activation complex between the leucine-responsive regulatory protein and DNA upstream of the gltBDF operon in Escherichia coli. J Mol Biol 270, 152–168.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.2008/018358-0
Loading
/content/journal/micro/10.1099/mic.0.2008/018358-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error