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. 1996 Aug 20;93(17):8841–8845. doi: 10.1073/pnas.93.17.8841

TnrA, a transcription factor required for global nitrogen regulation in Bacillus subtilis.

L V Wray Jr 1, A E Ferson 1, K Rohrer 1, S H Fisher 1
PMCID: PMC38555  PMID: 8799114

Abstract

Expression of the Bacillus subtilis nrgAB operon is derepressed during nitrogen-limited growth. We have identified a gene, tnrA, that is required for the activation of nrgAB expression under these growth conditions. Analysis of the DNA sequence of the tnrA gene revealed that it encodes a protein with sequence similarity to GlnR, the repressor of the B. subtilis glutamine synthetase operon. The tnrA mutant has a pleiotropic phenotype. Compared with wild-type cells, the tnrA mutant is impaired in its ability to utilize allantoin, gamma-aminobutyrate, isoleucine, nitrate, urea, and valine as nitrogen sources. During nitrogen-limited growth, transcription of the nrgAB, nasB, gabP, and ure genes is significantly reduced in the tnrA mutant compared with the levels seen in wild-type cells. In contrast, the level of glnRA expression is 4-fold higher in the, tnrA mutant than in wild-type cells during nitrogen restriction. The phenotype of the tnrA mutant indicates that a global nitrogen regulatory system is present in B. subtilis and that this system is distinct from the Ntr regulatory system found in enteric bacteria.

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Selected References

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

  1. Ahmed M., Borsch C. M., Taylor S. S., Vázquez-Laslop N., Neyfakh A. A. A protein that activates expression of a multidrug efflux transporter upon binding the transporter substrates. J Biol Chem. 1994 Nov 11;269(45):28506–28513. [PubMed] [Google Scholar]
  2. Amábile-Cuevas C. F., Demple B. Molecular characterization of the soxRS genes of Escherichia coli: two genes control a superoxide stress regulon. Nucleic Acids Res. 1991 Aug 25;19(16):4479–4484. doi: 10.1093/nar/19.16.4479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Atkinson M. R., Fisher S. H. Identification of genes and gene products whose expression is activated during nitrogen-limited growth in Bacillus subtilis. J Bacteriol. 1991 Jan;173(1):23–27. doi: 10.1128/jb.173.1.23-27.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Atkinson M. R., Wray L. V., Jr, Fisher S. H. Regulation of histidine and proline degradation enzymes by amino acid availability in Bacillus subtilis. J Bacteriol. 1990 Sep;172(9):4758–4765. doi: 10.1128/jb.172.9.4758-4765.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baumberg S., Harwood C. R. Carbon and nitrogen repression of arginine catabolic enzymes in Bacillus subtilis. J Bacteriol. 1979 Jan;137(1):189–196. doi: 10.1128/jb.137.1.189-196.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bender R. A. The role of the NAC protein in the nitrogen regulation of Klebsiella aerogenes. Mol Microbiol. 1991 Nov;5(11):2575–2580. doi: 10.1111/j.1365-2958.1991.tb01965.x. [DOI] [PubMed] [Google Scholar]
  7. Brown S. W., Sonenshein A. L. Autogenous regulation of the Bacillus subtilis glnRA operon. J Bacteriol. 1996 Apr;178(8):2450–2454. doi: 10.1128/jb.178.8.2450-2454.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dawes I. W., Mandelstam J. Sporulation of Bacillus subtilis in continuous culture. J Bacteriol. 1970 Sep;103(3):529–535. doi: 10.1128/jb.103.3.529-535.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dean D. R., Hoch J. A., Aronson A. I. Alteration of the Bacillus subtilis glutamine synthetase results in overproduction of the enzyme. J Bacteriol. 1977 Sep;131(3):981–987. doi: 10.1128/jb.131.3.981-987.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dodd I. B., Egan J. B. Improved detection of helix-turn-helix DNA-binding motifs in protein sequences. Nucleic Acids Res. 1990 Sep 11;18(17):5019–5026. doi: 10.1093/nar/18.17.5019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Débarbouillé M., Martin-Verstraete I., Kunst F., Rapoport G. The Bacillus subtilis sigL gene encodes an equivalent of sigma 54 from gram-negative bacteria. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9092–9096. doi: 10.1073/pnas.88.20.9092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Errington J., Vogt C. H. Isolation and characterization of mutations in the gene encoding an endogenous Bacillus subtilis beta-galactosidase and its regulator. J Bacteriol. 1990 Jan;172(1):488–490. doi: 10.1128/jb.172.1.488-490.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fisher S. H., Rosenkrantz M. S., Sonenshein A. L. Glutamine synthetase gene of Bacillus subtilis. Gene. 1984 Dec;32(3):427–438. doi: 10.1016/0378-1119(84)90018-0. [DOI] [PubMed] [Google Scholar]
  14. Fisher S. H., Sonenshein A. L. Bacillus subtilis glutamine synthetase mutants pleiotropically altered in glucose catabolite repression. J Bacteriol. 1984 Feb;157(2):612–621. doi: 10.1128/jb.157.2.612-621.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gutowski J. C., Schreier H. J. Interaction of the Bacillus subtilis glnRA repressor with operator and promoter sequences in vivo. J Bacteriol. 1992 Feb;174(3):671–681. doi: 10.1128/jb.174.3.671-681.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Holmes D. J., Caso J. L., Thompson C. J. Autogenous transcriptional activation of a thiostrepton-induced gene in Streptomyces lividans. EMBO J. 1993 Aug;12(8):3183–3191. doi: 10.1002/j.1460-2075.1993.tb05987.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Misra T. K., Brown N. L., Fritzinger D. C., Pridmore R. D., Barnes W. M., Haberstroh L., Silver S. Mercuric ion-resistance operons of plasmid R100 and transposon Tn501: the beginning of the operon including the regulatory region and the first two structural genes. Proc Natl Acad Sci U S A. 1984 Oct;81(19):5975–5979. doi: 10.1073/pnas.81.19.5975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Nakano M. M., Yang F., Hardin P., Zuber P. Nitrogen regulation of nasA and the nasB operon, which encode genes required for nitrate assimilation in Bacillus subtilis. J Bacteriol. 1995 Feb;177(3):573–579. doi: 10.1128/jb.177.3.573-579.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nakano Y., Kimura K. Purification and characterization of a repressor for the Bacillus cereus glnRA operon. J Biochem. 1991 Feb;109(2):223–228. [PubMed] [Google Scholar]
  20. 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]
  21. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schaeffer P., Millet J., Aubert J. P. Catabolic repression of bacterial sporulation. Proc Natl Acad Sci U S A. 1965 Sep;54(3):704–711. doi: 10.1073/pnas.54.3.704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schreier H. J., Brown S. W., Hirschi K. D., Nomellini J. F., Sonenshein A. L. Regulation of Bacillus subtilis glutamine synthetase gene expression by the product of the glnR gene. J Mol Biol. 1989 Nov 5;210(1):51–63. doi: 10.1016/0022-2836(89)90290-8. [DOI] [PubMed] [Google Scholar]
  24. Schreier H. J., Fisher S. H., Sonenshein A. L. Regulation of expression from the glnA promoter of Bacillus subtilis requires the glnA gene product. Proc Natl Acad Sci U S A. 1985 May;82(10):3375–3379. doi: 10.1073/pnas.82.10.3375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schreier H. J., Rostkowski C. A. Bacillus subtilis glnR mutants defective in regulation. Gene. 1995 Aug 8;161(1):51–56. doi: 10.1016/0378-1119(95)00238-2. [DOI] [PubMed] [Google Scholar]
  26. Shewchuk L. M., Helmann J. D., Ross W., Park S. J., Summers A. O., Walsh C. T. Transcriptional switching by the MerR protein: activation and repression mutants implicate distinct DNA and mercury(II) binding domains. Biochemistry. 1989 Mar 7;28(5):2340–2344. doi: 10.1021/bi00431a053. [DOI] [PubMed] [Google Scholar]
  27. Sonenshein A. L., Cami B., Brevet J., Cote R. Isolation and characterization of rifampin-resistant and streptolydigin-resistant mutants of Bacillus subtilis with altered sporulation properties. J Bacteriol. 1974 Oct;120(1):253–265. doi: 10.1128/jb.120.1.253-265.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Strauch M. A., Aronson A. I., Brown S. W., Schreier H. J., Sonenhein A. L. Sequence of the Bacillus subtilis glutamine synthetase gene region. Gene. 1988 Nov 30;71(2):257–265. doi: 10.1016/0378-1119(88)90042-x. [DOI] [PubMed] [Google Scholar]
  29. Sun D. X., Setlow P. Cloning, nucleotide sequence, and expression of the Bacillus subtilis ans operon, which codes for L-asparaginase and L-aspartase. J Bacteriol. 1991 Jun;173(12):3831–3845. doi: 10.1128/jb.173.12.3831-3845.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Vega-Palas M. A., Flores E., Herrero A. NtcA, a global nitrogen regulator from the cyanobacterium Synechococcus that belongs to the Crp family of bacterial regulators. Mol Microbiol. 1992 Jul;6(13):1853–1859. doi: 10.1111/j.1365-2958.1992.tb01357.x. [DOI] [PubMed] [Google Scholar]
  31. Wray L. V., Jr, Atkinson M. R., Fisher S. H. The nitrogen-regulated Bacillus subtilis nrgAB operon encodes a membrane protein and a protein highly similar to the Escherichia coli glnB-encoded PII protein. J Bacteriol. 1994 Jan;176(1):108–114. doi: 10.1128/jb.176.1.108-114.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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