Summary
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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ames G Ferro-Luzzi (1986) Bacterial periplasmic transport systems: structure, mechanism, and evolution. Annu Rev Biochem 55:397–425
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 4:539–547
Ausubel FM (1984) Regulation of nitrogen fixation genes. Cell 37:5–6
Beynon J, Cannon M, Buchanan-Wollaston V, Cannon F (1983) The nif promoters of Klebsiella pneumoniae have a characteristic primary structure. Cell 34:665–671
Brown ES, Ausubel FM (1984) Mutation affecting regulation of the Klebsiella pneumoniae nif RH (nitrogenase reductase) promoter. J Bacteriol 157:143–147
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 13:7621–7638
Buck M, Miller S, Drummond M, Dixon R (1986) Upstream activator sequences are present in the promoters of nitrogen fixation genes. Nature 320:374–378
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 183:289–297
Dixon RA (1984) The genetic complexity of nitrogen fixation. J Gen Microbiol 130:2745–2755
Drummond M, Clements J, Merrick C, Dixon R (1983) Positive control and autogenous regulation of the nifLA promoter in Klebsiella pneumoniae. Nature 301:302–307
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 100:215–219
Higgins CF, Ames G Ferro-Luzzi (1982) Regulatory regions of two transport operons under nitrogen control: nucleotide sequences. Proc Natl Acad Sci USA 79:1083–1087
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 82:7525–7529
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 82:8453–8457
Kunkel TA (1985) Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc Natl Acad Sci USA 82:488–492
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 138:218–234
Magasanik B (1982) Genetic control of nitrogen assimilation in bacteria. Annu Rev Genet 16:135–168
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
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 383–415
Merrick MJ (1982) A new model for nitrogen control. Nature 297:362–363
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 2:39–44
Messing J (1983) New M13 vectors for cloning. Methods Enzymol 101:20–78
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
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 83:5909–5913
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 78:2213–2217
Ow DW, Ausubel FM (1983) Regulation of nitrogen metabolism genes by nifA gene product in Klebsiella pneumoniae. Nature 301:307–313
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 161:868–874
Raibaud O, Schwartz M (1984) Positive control of transcription initiation in bacteria. Annu Rev Genet 18:173–206
Reitzer LJ, Magasanik B (1985) Expression of glnA in Escherichia coli is regulated at tandem promoters. Proc Natl Acad Sci USA 82:1979–1983
Reitzer LJ, Magasanik B (1986) Transcription of glnA in Escherichia coli is stimulated by activator bound to sites far from the promoter. Cell 45:785–792
Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet 13:319–353
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 150:221–230
Sanger F, Nicklen S, Coulson A (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5476
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 209:403–407
Stern MJ, Higgins CF, Ames G Ferro-Luzzi (1984) Isolation and characterization of lac fusions to two nitrogen-regulated promoters. Mol Gen Genet 195:219–227
Tumer NE, Robinson SJ, Haselkorn R (1983) Different promoters for the Anabaena glutamine synthetase gene during growth using molecular or fixed nitrogen. Nature 306:337–342
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 160:379–384
Zoller MJ, Smith M (1983) Oligonucleotide directed mutagenesis of DNA fragments cloned into M13 vectors. Methods Enzymol 100:468–500
Author information
Authors and Affiliations
Additional information
Communicated by J. Lengeler
Rights and permissions
About this article
Cite this article
Schmitz, G., Nikaido, K. & Ames, G.FL. Regulation of a transport operon promoter in Salmonella typhimurium: Identification of sites essential for nitrogen regulation. Mol Gen Genet 215, 107–117 (1988). https://doi.org/10.1007/BF00331311
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00331311