Molecular and General Genetics MGG

, Volume 215, Issue 1, pp 107–117 | Cite as

Regulation of a transport operon promoter in Salmonella typhimurium: Identification of sites essential for nitrogen regulation

  • Gudrun Schmitz
  • Kishiko Nikaido
  • Giovanna Ferro-Luzzi Ames


The promoter of nitrogen-regulated transport, argTr, has been mutationally altered in order to determine the features that are essential for its response to nitrogen availability. Deletions of all sequences upstream of position-44 or downstream of position +2 had no effect no nitrogen regulation of argTr. These deletions define a small region of 44 bp where all necessary features for nitrogen regulation are located. This region includes for nitrogen regulation are located. This region includes sequences highly homologous to the nif consensus promoter. Alteration of this particular sequence caused drastic changes in the response to changes of nitrogen availability, thus indicating that they are directly involved in regulation. This implies that the NtrC protein must also act within this small region of the promoter. The data are discussed in terms of current-hypotheses concerning nitrogen regulation. In addition, we have shown 1. that carbon regulation at this promoter must occur at a site upstream from the nitrogen promoter; 2. that nifA can replace ntrC in the regulation of argTr.

Key words

Carbon regulation Nitrogen regulation ntrC activation Transport 


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  1. Ames G Ferro-Luzzi (1986) Bacterial periplasmic transport systems: structure, mechanism, and evolution. Annu Rev Biochem 55:397–425Google Scholar
  2. Ames G Ferro-Luzzi, Nikaido K (1985) Nitrogen regulation in Salmonella typhimurium. Identification of an ntrC protein-binding site and definition of a consensus binding sequence. EMBO J 4:539–547Google Scholar
  3. Ausubel FM (1984) Regulation of nitrogen fixation genes. Cell 37:5–6Google Scholar
  4. Beynon J, Cannon M, Buchanan-Wollaston V, Cannon F (1983) The nif promoters of Klebsiella pneumoniae have a characteristic primary structure. Cell 34:665–671Google Scholar
  5. Brown ES, Ausubel FM (1984) Mutation affecting regulation of the Klebsiella pneumoniae nif RH (nitrogenase reductase) promoter. J Bacteriol 157:143–147Google Scholar
  6. Buck M, Khan H, Dixon R (1985) Site-directed mutagenesis of the Klebsiella pneumoniae nifL and nifH promoters and in vivo analysis of promoter activity. Nucleic Acids Res 13:7621–7638Google Scholar
  7. Buck M, Miller S, Drummond M, Dixon R (1986) Upstream activator sequences are present in the promoters of nitrogen fixation genes. Nature 320:374–378Google Scholar
  8. deBruijn FJ, Ausubel FM (1981) The cloning and transposon Tn5 mutagenesis of the glnA region of Klebsiella pneumoniae: identification of glnR, a gene involved in the regulation of the nif and hut operons. Mol Gen Genet 183:289–297Google Scholar
  9. Dixon RA (1984) The genetic complexity of nitrogen fixation. J Gen Microbiol 130:2745–2755Google Scholar
  10. Drummond M, Clements J, Merrick C, Dixon R (1983) Positive control and autogenous regulation of the nifLA promoter in Klebsiella pneumoniae. Nature 301:302–307Google Scholar
  11. Gutnick D, Calvo JM, Klopotowski T, Ames BN (1969) Compounds which serve as the sole source of carbon or nitrogen for Salmonella typhimurium LT-2. J Bacteriol 100:215–219Google Scholar
  12. Higgins CF, Ames G Ferro-Luzzi (1982) Regulatory regions of two transport operons under nitrogen control: nucleotide sequences. Proc Natl Acad Sci USA 79:1083–1087Google Scholar
  13. Hirschman J, Wong P-K, Sei K, Keener J, Kustu S (1985) Products of nitrogen regulatory genes ntrA and ntrC of enteric bacteria activate glnA transcription in vitro: evidence that the ntrA product is a σ factor. Proc Natl Acad Sci USA 82:7525–7529Google Scholar
  14. Hunt TP, Magasanik B (1985) Transcription of glnA by purified Escherichia coli components: core RNA polymerase and the products of glnF, glnG, and glnL. Proc Natl Acad Sci USA 82:8453–8457Google Scholar
  15. Kunkel TA (1985) Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci USA 82:488–492Google Scholar
  16. Kustu SG, McFarland NC, Hui SP, Esmon B, Ames G Ferro-Luzzi (1979b) Nitrogen control in Salmonella typhimurium: co-regulation of synthesis of glutamine synthetase and amino acid transport systems. J Bacteriol 138:218–234Google Scholar
  17. Magasanik B (1982) Genetic control of nitrogen assimilation in bacteria. Annu Rev Genet 16:135–168Google Scholar
  18. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
  19. McKenney K, Shimatahe H, Court D, Schmeissner U, Brady C, Rosenberg M (1981) In: Chirikijan JC, Papas TS (eds) Gene amplification and analysis, vol II. Elsevier/North Holland, New York, pp 383–415Google Scholar
  20. Merrick MJ (1982) A new model for nitrogen control. Nature 297:362–363Google Scholar
  21. Merrick MJ (1983) Nitrogen control of the NIF regulon in Klebsiella pneumoniae involvement of the ntrA gene and analogies between ntrC and nifA. EMBO J 2:39–44Google Scholar
  22. Messing J (1983) New M13 vectors for cloning. Methods Enzymol 101:20–78Google Scholar
  23. Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
  24. Ninfa AJ, Magasanik B (1986) Covalent modification of the glnG product, NRI, by the glnL product, NRII, regulates the transcription of the glnALG operon in Escherichia coli. Proc Natl Acad Sci USA 83:5909–5913Google Scholar
  25. O'Neill MC, Amass K, De Crombrugghe B (1981) Molecular model of the DNA interaction site for the cyclic AMP receptor protein. Proc Natl Acad Sci USA 78:2213–2217Google Scholar
  26. Ow DW, Ausubel FM (1983) Regulation of nitrogen metabolism genes by nifA gene product in Klebsiella pneumoniae. Nature 301:307–313Google Scholar
  27. Ow DW, Xiong Y, Gu Q, Shen SC (1985) Mutational analysis of the Klebsiella pneumoniae nitrogenase promoter; sequences essential for positive control by nifA and ntrC (glnG) products. J Bacteriol 161:868–874Google Scholar
  28. Raibaud O, Schwartz M (1984) Positive control of transcription initiation in bacteria. Annu Rev Genet 18:173–206Google Scholar
  29. Reitzer LJ, Magasanik B (1985) Expression of glnA in Escherichia coli is regulated at tandem promoters. Proc Natl Acad Sci USA 82:1979–1983Google Scholar
  30. Reitzer LJ, Magasanik B (1986) Transcription of glnA in Escherichia coli is stimulated by activator bound to sites far from the promoter. Cell 45:785–792Google Scholar
  31. Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet 13:319–353Google Scholar
  32. Rothman N, Rothstein D, Foor F, Magasanik KB (1982) Role of glnA-linked genes in regulation of glutamine synthetase and histidase formation in Klebsiella aerogenes. J Bacteriol 150:221–230Google Scholar
  33. Sanger F, Nicklen S, Coulson A (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5476Google Scholar
  34. Schmitz G, Dürre P, Mullenbach G, Ames G Ferro-Luzzi (1987) Nitrogen regulation of transport operons: analysis of promoters argTr and dhuA. Mol Gen Genet 209:403–407Google Scholar
  35. Stern MJ, Higgins CF, Ames G Ferro-Luzzi (1984) Isolation and characterization of lac fusions to two nitrogen-regulated promoters. Mol Gen Genet 195:219–227Google Scholar
  36. Tumer NE, Robinson SJ, Haselkorn R (1983) Different promoters for the Anabaena glutamine synthetase gene during growth using molecular or fixed nitrogen. Nature 306:337–342Google Scholar
  37. Ueno-Nishio S, Mango S, Reitzer LJ, Magasanik B (1984) Identification and regulation of glnL operator-promoter of the complex glnALG operon of Escherichia coli. J Bacteriol 160:379–384Google Scholar
  38. Zoller MJ, Smith M (1983) Oligonucleotide directed mutagenesis of DNA fragments cloned into M13 vectors. Methods Enzymol 100:468–500Google Scholar

Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • Gudrun Schmitz
    • 1
  • Kishiko Nikaido
    • 2
  • Giovanna Ferro-Luzzi Ames
    • 2
  1. 1.Boehringer Mannheim GmbHPenzbergGermany
  2. 2.Department of BiochemistryUniversity of CaliforniaBerkeleyUSA

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