This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Wray, L. V.
Right arrow Articles by Fisher, S. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Wray, L. V., Jr.
Right arrow Articles by Fisher, S. H.

 Previous Article  |  Next Article 

J Bacteriol, June 1998, p. 2943-2949, Vol. 180, No. 11
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Mutational Analysis of the TnrA-Binding Sites in the Bacillus subtilis nrgAB and gabP Promoter Regions

Lewis V. Wray Jr., Jill M. Zalieckas, Amy E. Ferson, and Susan H. Fisher*

Department of Microbiology, Boston University School of Medicine, Boston, Massachusetts 02118

Received 26 January 1998/Accepted 6 April 1998

Transcription of the Bacillus subtilis nrgAB promoter is activated during nitrogen-limited growth by the TnrA protein. A common inverted repeat, TGTNAN7TNACA (TnrA site), is centered 49 to 51 bp upstream of the transcriptional start sites for the TnrA-regulated nrgAB, gabP P2, and nas promoters. Oligonucleotide-directed mutagenesis of the nrgAB promoter region showed that conserved nucleotides within the TnrA site, the A+T-rich region between the two TnrA half-sites, and an upstream A tract are all required for high-level activation of nrgAB expression. Mutations that alter the relative distance between the two half-sites of the nrgAB TnrA site abolish nitrogen regulation of nrgAB expression. Spacer mutations that change the relative distance between the TnrA site and -35 region of the nrgAB promoter reveal that activation of nrgAB expression occurs only when the TnrA site is located 49 to 51 bp upstream of the transcriptional start site. Mutational analysis of the conserved nucleotides in the gabP P2 TnrA site showed that this sequence is also required for nitrogen-regulated gabP P2 expression. The TnrA protein, expressed in an overproducing Escherichia coli strain, had a 625-fold-higher affinity for the wild-type nrgAB promoter DNA than for a mutated nrgAB promoter DNA fragment that is unable to activate nrgAB expression in vivo. These results indicate that the proposed TnrA site functions as the binding site for the TnrA protein. TnrA was found to activate nrgAB expression during late exponential growth in nutrient sporulation medium containing glucose, suggesting that cells become nitrogen limited during growth in this medium.

* Corresponding author. Mailing address: Department of Microbiology, Boston University School of Medicine, 715 Albany St., Boston, MA 02118. Phone: (617) 638-5498. Fax: (617) 638-4286. E-mail: shfisher{at}bu.edu.

J Bacteriol, June 1998, p. 2943-2949, Vol. 180, No. 11
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Fisher, S. H., Wray, L. V. Jr. (2009). Novel trans-Acting Bacillus subtilis glnA Mutations That Derepress glnRA Expression. J. Bacteriol. 191: 2485-2492 [Abstract] [Full Text]  
  • Kayumov, A., Heinrich, A., Sharipova, M., Iljinskaya, O., Forchhammer, K. (2008). Inactivation of the general transcription factor TnrA in Bacillus subtilis by proteolysis. Microbiology 154: 2348-2355 [Abstract] [Full Text]  
  • Yasumura, A., Abe, S., Tanaka, T. (2008). Involvement of Nitrogen Regulation in Bacillus subtilis degU Expression. J. Bacteriol. 190: 5162-5171 [Abstract] [Full Text]  
  • Kloosterman, T. G., Hendriksen, W. T., Bijlsma, J. J. E., Bootsma, H. J., van Hijum, S. A. F. T., Kok, J., Hermans, P. W. M., Kuipers, O. P. (2006). Regulation of Glutamine and Glutamate Metabolism by GlnR and GlnA in Streptococcus pneumoniae. J. Biol. Chem. 281: 25097-25109 [Abstract] [Full Text]  
  • Fisher, S. H., Wray, L. V. Jr (2006). Feedback-Resistant Mutations in Bacillus subtilis Glutamine Synthetase Are Clustered in the Active Site.. J. Bacteriol. 188: 5966-5974 [Abstract] [Full Text]  
  • Zalieckas, J. M., Wray, L. V. Jr., Fisher, S. H. (2006). Cross-Regulation of the Bacillus subtilis glnRA and tnrA Genes Provides Evidence for DNA Binding Site Discrimination by GlnR and TnrA.. J. Bacteriol. 188: 2578-2585 [Abstract] [Full Text]  
  • Brandenburg, J. L., Wray, L. V. Jr., Beier, L., Jarmer, H., Saxild, H. H., Fisher, S. H. (2002). Roles of PucR, GlnR, and TnrA in Regulating Expression of the Bacillus subtilisure P3 Promoter. J. Bacteriol. 184: 6060-6064 [Abstract] [Full Text]  
  • Beier, L., Nygaard, P., Jarmer, H., Saxild, H. H. (2002). Transcription Analysis of the Bacillus subtilis PucR Regulon and Identification of a cis-Acting Sequence Required for PucR-Regulated Expression of Genes Involved in Purine Catabolism. J. Bacteriol. 184: 3232-3241 [Abstract] [Full Text]  
  • Fisher, S. H., Wray, L. V. Jr. (2002). Bacillus subtilis 168 Contains Two Differentially Regulated Genes Encoding L-Asparaginase. J. Bacteriol. 184: 2148-2154 [Abstract] [Full Text]  
  • Arcondeguy, T., Jack, R., Merrick, M. (2001). PII Signal Transduction Proteins, Pivotal Players in Microbial Nitrogen Control. Microbiol. Mol. Biol. Rev. 65: 80-105 [Abstract] [Full Text]  
  • Belitsky, B. R., Wray, L. V. Jr., Fisher, S. H., Bohannon, D. E., Sonenshein, A. L. (2000). Role of TnrA in Nitrogen Source-Dependent Repression of Bacillus subtilis Glutamate Synthase Gene Expression. J. Bacteriol. 182: 5939-5947 [Abstract] [Full Text]  
  • Shin, B.-S., Choi, S.-K., Smith, I., Park, S.-H. (2000). Analysis of tnrA Alleles Which Result in a Glucose-Resistant Sporulation Phenotype in Bacillus subtilis. J. Bacteriol. 182: 5009-5012 [Abstract] [Full Text]  
  • Robichon, D., Arnaud, M., Gardan, R., Pragai, Z., O'Reilly, M., Rapoport, G., Débarbouillé, M. (2000). Expression of a New Operon from Bacillus subtilis, ykzB-ykoL, under the Control of the TnrA and PhoP-PhoR Global Regulators. J. Bacteriol. 182: 1226-1231 [Abstract] [Full Text]  
  • Zalieckas, J. M., Wray, L. V. Jr., Fisher, S. H. (1999). trans-Acting Factors Affecting Carbon Catabolite Repression of the hut Operon in Bacillus subtilis. J. Bacteriol. 181: 2883-2888 [Abstract] [Full Text]  
  • Nakano, M. M., Hoffmann, T., Zhu, Y., Jahn, D. (1998). Nitrogen and Oxygen Regulation of Bacillus subtilis nasDEF Encoding NADH-Dependent Nitrite Reductase by TnrA and ResDE. J. Bacteriol. 180: 5344-5350 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»