Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Feb;174(4):1109–1118. doi: 10.1128/jb.174.4.1109-1118.1992

Characterization of the regulon controlled by the leucine-responsive regulatory protein in Escherichia coli.

B R Ernsting 1, M R Atkinson 1, A J Ninfa 1, R G Matthews 1
PMCID: PMC206403  PMID: 1346534

Abstract

The leucine-responsive regulatory protein (Lrp) has been shown to regulate, either positively or negatively, the transcription of several Escherichia coli genes in response to leucine. We have used two-dimensional gel electrophoresis to analyze the patterns of polypeptide expression in isogenic lrp+ and lrp mutant strains in the presence or absence of leucine. The absence of a functional Lrp protein alters the expression of at least 30 polypeptides. The expression of the majority of these polypeptides is not affected by the presence or absence of 10 mM exogenous leucine. Outer membrane porins OmpC and OmpF, glutamine synthetase (GlnA), the small subunit of glutamate synthase (GltD), lysyl-tRNA synthetase form II (LysU), a high-affinity periplasmic binding protein specific for branched-chain amino acids (LivJ), W protein, and the enzymes of the pathway converting threonine to glycine, namely, threonine dehydrogenase (Tdh) and 2-amino-3-ketobutyrate coenzyme A ligase (Kbl), were identified as members of the Lrp regulon by electrophoretic analysis. We have shown that Lrp is a positive regulator of glutamate synthase and glutamine synthetase and that exogenous leucine has little or no effect on the expression of these proteins. In strains carrying a glnL deletion and in strains carrying the glnL2302 allele, which directs the synthesis of a GlnL protein that is constitutively active, expression of glutamine synthetase is no longer regulated by Lrp, demonstrating that the effect of Lrp on glutamine synthetase levels is indirect and requires an intact glnL gene. lrp::Tn10 strains grow poorly when arginine or ornithine is present as the sole nitrogen source in the medium. On the bases of present studies and previous research, we propose that Lrp is involved in the adaptation of E. coli cells to major shifts in environment, such as those which occur when E. coli leaves the intestinal tract of its animal host. Several genes required for amino acid and peptide transport and catabolism are negatively regulated by Lrp, and other genes required for amino acid biosynthesis and ammonia assimilation in a nitrogen-poor environment are positively regulated by Lrp.

Full text

PDF
1109

Images in this article

Selected References

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

  1. Backman K., Chen Y. M., Magasanik B. Physical and genetic characterization of the glnA--glnG region of the Escherichia coli chromosome. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3743–3747. doi: 10.1073/pnas.78.6.3743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Blumenthal R. M., Reeh S., Pedersen S. Regulation of transcription factor rho and the alpha subunit of RNA polymerase in Escherichia coli B/r. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2285–2288. doi: 10.1073/pnas.73.7.2285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bower S., Zalkin H. Chemical modification and ligand binding studies with Escherichia coli glutamate synthase. Biochemistry. 1983 Mar 29;22(7):1613–1620. doi: 10.1021/bi00276a014. [DOI] [PubMed] [Google Scholar]
  4. Boylan S. A., Dekker E. E. L-threonine dehydrogenase. Purification and properties of the homogeneous enzyme from Escherichia coli K-12. J Biol Chem. 1981 Feb 25;256(4):1809–1815. [PubMed] [Google Scholar]
  5. Brenchley J. E., Prival M. J., Magasanik B. Regulation of the synthesis of enzymes responsible for glutamate formation in Klebsiella aerogenes. J Biol Chem. 1973 Sep 10;248(17):6122–6128. [PubMed] [Google Scholar]
  6. Bueno R., Pahel G., Magasanik B. Role of glnB and glnD gene products in regulation of the glnALG operon of Escherichia coli. J Bacteriol. 1985 Nov;164(2):816–822. doi: 10.1128/jb.164.2.816-822.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen Y. M., Backman K., Magasanik B. Characterization of a gene, glnL, the product of which is involved in the regulation of nitrogen utilization in Escherichia coli. J Bacteriol. 1982 Apr;150(1):214–220. doi: 10.1128/jb.150.1.214-220.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ganoza M. C., Cunningham C., Green R. M. Isolation and point of action of a factor from Escherichia coli required to reconstruct translation. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1648–1652. doi: 10.1073/pnas.82.6.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Garciarrubio A., Lozoya E., Covarrubias A., Bolivar F. Structural organization of the genes that encode two glutamate synthase subunits of Escherichia coli. Gene. 1983 Dec;26(2-3):165–170. doi: 10.1016/0378-1119(83)90186-5. [DOI] [PubMed] [Google Scholar]
  10. Garrett S., Taylor R. K., Silhavy T. J. Isolation and characterization of chain-terminating nonsense mutations in a porin regulator gene, envZ. J Bacteriol. 1983 Oct;156(1):62–69. doi: 10.1128/jb.156.1.62-69.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gollop N., Tavori H., Barak Z. Acetohydroxy acid synthase is a target for leucine containing peptide toxicity in Escherichia coli. J Bacteriol. 1982 Jan;149(1):387–390. doi: 10.1128/jb.149.1.387-390.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Himmelfarb H. J., Pearlberg J., Last D. H., Ptashne M. GAL11P: a yeast mutation that potentiates the effect of weak GAL4-derived activators. Cell. 1990 Dec 21;63(6):1299–1309. doi: 10.1016/0092-8674(90)90425-e. [DOI] [PubMed] [Google Scholar]
  13. Hirshfield I. N., Bloch P. L., Van Bogelen R. A., Neidhardt F. C. Multiple forms of lysyl-transfer ribonucleic acid synthetase in Escherichia coli. J Bacteriol. 1981 Apr;146(1):345–351. doi: 10.1128/jb.146.1.345-351.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. Matthews R. G., Neidhardt F. C. Abnormal induction of heat shock proteins in an Escherichia coli mutant deficient in adenosylmethionine synthetase activity. J Bacteriol. 1988 Apr;170(4):1582–1588. doi: 10.1128/jb.170.4.1582-1588.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mäntsälä P., Zalkin H. Glutamate synthase. Properties of the glutamine-dependent activity. J Biol Chem. 1976 Jun 10;251(11):3294–3299. [PubMed] [Google Scholar]
  17. Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  19. Pahel G., Rothstein D. M., Magasanik B. Complex glnA-glnL-glnG operon of Escherichia coli. J Bacteriol. 1982 Apr;150(1):202–213. doi: 10.1128/jb.150.1.202-213.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pahel G., Zelenetz A. D., Tyler B. M. gltB gene and regulation of nitrogen metabolism by glutamine synthetase in Escherichia coli. J Bacteriol. 1978 Jan;133(1):139–148. doi: 10.1128/jb.133.1.139-148.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Quay S. C., Dick T. E., Oxender D. L. Role of transport systems in amino acid metabolism: leucine toxicity and the branched-chain amino acid transport systems. J Bacteriol. 1977 Mar;129(3):1257–1265. doi: 10.1128/jb.129.3.1257-1265.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Ricca E., Aker D. A., Calvo J. M. A protein that binds to the regulatory region of the Escherichia coli ilvIH operon. J Bacteriol. 1989 Mar;171(3):1658–1664. doi: 10.1128/jb.171.3.1658-1664.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stadtman E. R., Ginsburg A., Ciardi J. E., Yeh J., Hennig S. B., Shapiro B. M. Multiple molecular forms of glutamine synthetase produced by enzyme catalyzed adenylation and deadenylylation reactions. Adv Enzyme Regul. 1970;8:99–118. doi: 10.1016/0065-2571(70)90011-7. [DOI] [PubMed] [Google Scholar]
  26. Tabata S., Higashitani A., Takanami M., Akiyama K., Kohara Y., Nishimura Y., Nishimura A., Yasuda S., Hirota Y. Construction of an ordered cosmid collection of the Escherichia coli K-12 W3110 chromosome. J Bacteriol. 1989 Feb;171(2):1214–1218. doi: 10.1128/jb.171.2.1214-1218.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. VanBogelen R. A., Neidhardt F. C. The gene-protein database of Escherichia coli: edition 4. Electrophoresis. 1991 Nov;12(11):955–994. doi: 10.1002/elps.1150121114. [DOI] [PubMed] [Google Scholar]
  29. Wanner B. L., Kodaira R., Neidhardt F. C. Physiological regulation of a decontrolled lac operon. J Bacteriol. 1977 Apr;130(1):212–222. doi: 10.1128/jb.130.1.212-222.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]

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

RESOURCES