Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1983 Jul;80(13):4030–4034. doi: 10.1073/pnas.80.13.4030

Activation of Klebsiella pneumoniae and Rhizobium meliloti nitrogenase promoters by gln (ntr) regulatory proteins.

V Sundaresan, D W Ow, F M Ausubel
PMCID: PMC394194  PMID: 6306658

Abstract

We have studied the expression, in different Escherichia coli gln (ntr) mutants, of fusions (constructed in vitro) of the nifHDK (nitrogenase) promoters from Klebsiella pneumoniae and Rhizobium meliloti to E. coli lacZ. Derepression of the K. pneumoniae nifH::lacZ fusion requires the glnF (ntrA) gene product in addition to the K. pneumoniae nifA gene product, indicating that regulation of the K. pneumoniae nif genes is more closely integrated with the overall nitrogen control system than previously demonstrated. Derepression of the R. meliloti nifH::lacZ fusion in E. coli by the K. pneumoniae nifA gene product (which we had previously shown) exhibits the same requirement for glnF. Derepression of the R. meliloti nifH::lacZ fusion, but not the K. pneumoniae nifH::lacZ fusion, can be mediated by the glnG (ntrC) gene product, suggesting that the gln regulatory genes might directly regulate the symbiotic nitrogen fixation genes in Rhizobium.

Full text

PDF
4030

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. Buchanan-Wollaston V., Cannon M. C., Beynon J. L., Cannon F. C. Role of the nifA gene product in the regulation of nif expression in Klebsiella pneumoniae. Nature. 1981 Dec 24;294(5843):776–778. doi: 10.1038/294776a0. [DOI] [PubMed] [Google Scholar]
  3. Casadaban M. J., Chou J., Cohen S. N. In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals. J Bacteriol. 1980 Aug;143(2):971–980. doi: 10.1128/jb.143.2.971-980.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Espin G., Alvarez-Morales A., Merrick M. Complementation analysis of glnA-linked mutations which affect nitrogen fixation in Klebsiella pneumoniae. Mol Gen Genet. 1981;184(2):213–217. doi: 10.1007/BF00272907. [DOI] [PubMed] [Google Scholar]
  6. Foor F., Cedergren R. J., Streicher S. L., Rhee S. G., Magasanik B. Glutamine synthetase of Klebsiella aerogenes: properties of glnD mutants lacking uridylyltransferase. J Bacteriol. 1978 May;134(2):562–568. doi: 10.1128/jb.134.2.562-568.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Foor F., Reuveny Z., Magasanik B. Regulation of the synthesis of glutamine synthetase by the PII protein in Klebsiella aerogenes. Proc Natl Acad Sci U S A. 1980 May;77(5):2636–2640. doi: 10.1073/pnas.77.5.2636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kennedy C., Robson R. L. Activation of nif gene expression in Azotobacter by the nifA gene product of Klebsiella pneumoniae. Nature. 1983 Feb 17;301(5901):626–628. doi: 10.1038/301626a0. [DOI] [PubMed] [Google Scholar]
  9. Kustu S., Burton D., Garcia E., McCarter L., McFarland N. Nitrogen control in Salmonella: regulation by the glnR and glnF gene products. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4576–4580. doi: 10.1073/pnas.76.9.4576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ludwig R. A. Regulation of Rhizobium nitrogen fixation by the unadenylylated glutamine synthetase I system. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5817–5821. doi: 10.1073/pnas.77.10.5817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. McFarland N., McCarter L., Artz S., Kustu S. Nitrogen regulatory locus "glnR" of enteric bacteria is composed of cistrons ntrB and ntrC: identification of their protein products. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2135–2139. doi: 10.1073/pnas.78.4.2135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Merrick M. J. A new model for nitrogen control. Nature. 1982 Jun 3;297(5865):362–363. doi: 10.1038/297362a0. [DOI] [PubMed] [Google Scholar]
  13. Merrick M. J. Nitrogen control of the nif regulon in Klebsiella pneumoniae: involvement of the ntrA gene and analogies between ntrC and nifA. EMBO J. 1983;2(1):39–44. doi: 10.1002/j.1460-2075.1983.tb01377.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ow D. W., Ausubel F. M. Recombinant P4 bacteriophages propagate as viable lytic phages or as autonomous plasmids in Klebsiella pneumoniae. Mol Gen Genet. 1980;180(1):165–175. doi: 10.1007/BF00267366. [DOI] [PubMed] [Google Scholar]
  15. Ow D. W., Ausubel F. M. Regulation of nitrogen metabolism genes by nifA gene product in Klebsiella pneumoniae. Nature. 1983 Jan 27;301(5898):307–313. doi: 10.1038/301307a0. [DOI] [PubMed] [Google Scholar]
  16. Ow D. W., Sundaresan V., Rothstein D. M., Brown S. E., Ausubel F. M. Promoters regulated by the glnG (ntrC) and nifA gene products share a heptameric consensus sequence in the -15 region. Proc Natl Acad Sci U S A. 1983 May;80(9):2524–2528. doi: 10.1073/pnas.80.9.2524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Roberts G. P., Brill W. J. Genetics and regulation of nitrogen fixation. Annu Rev Microbiol. 1981;35:207–235. doi: 10.1146/annurev.mi.35.100181.001231. [DOI] [PubMed] [Google Scholar]
  19. Ruvkun G. B., Ausubel F. M. Interspecies homology of nitrogenase genes. Proc Natl Acad Sci U S A. 1980 Jan;77(1):191–195. doi: 10.1073/pnas.77.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ruvkun G. B., Sundaresan V., Ausubel F. M. Directed transposon Tn5 mutagenesis and complementation analysis of Rhizobium meliloti symbiotic nitrogen fixation genes. Cell. 1982 Jun;29(2):551–559. doi: 10.1016/0092-8674(82)90171-4. [DOI] [PubMed] [Google Scholar]
  21. Sibold L., Elmerich C. Constitutive expression of nitrogen fixation (nif) genes of Klebsiella pneumoniae due to a DNA duplication. EMBO J. 1982;1(12):1551–1558. doi: 10.1002/j.1460-2075.1982.tb01354.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sundaresan V., Ausubel F. M. Nucleotide sequence of the gene coding for the nitrogenase iron protein from Klebsiella pneumoniae. J Biol Chem. 1981 Mar 25;256(6):2808–2812. [PubMed] [Google Scholar]
  23. Sundaresan V., Jones J. D., Ow D. W., Ausubel F. M. Klebsiella pneumoniae nifA product activates the Rhizobium meliloti nitrogenase promoter. Nature. 1983 Feb 24;301(5902):728–732. doi: 10.1038/301728a0. [DOI] [PubMed] [Google Scholar]
  24. Török I., Kondorosi A. Nucleotide sequence of the R.meliloti nitrogenase reductase (nifH) gene. Nucleic Acids Res. 1981 Nov 11;9(21):5711–5723. doi: 10.1093/nar/9.21.5711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. de Bruijn F. J., Ausubel F. M. 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. 1981;183(2):289–297. doi: 10.1007/BF00270631. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES