Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Sep;177(18):5322–5326. doi: 10.1128/jb.177.18.5322-5326.1995

Effect of an ntrBC mutation on the posttranslational regulation of nitrogenase activity in Rhodospirillum rubrum.

Y Zhang 1, A D Cummings 1, R H Burris 1, P W Ludden 1, G P Roberts 1
PMCID: PMC177326  PMID: 7665521

Abstract

Homologs of ntrB and ntrC genes from Rhodospirillum rubrum were cloned and sequenced. A mutant lacking ntrBC was constructed, and this mutant has normal nitrogenase activity under nif-derepressing conditions, indicating that ntrBC are not necessary for the expression of the nif genes in R. rubrum. However, the post-translational regulation of nitrogenase activity by ADP-ribosylation in response to NH4+ was partially abolished in this mutant. More surprisingly, the regulation of nitrogenase activity in response to darkness was also affected, suggesting a physiological link between the ntr system and energy signal transduction in R. rubrum. The expression of glutamine synthetase, as well as its posttranslational regulation, was also altered in this ntrBC mutant.

Full Text

The Full Text of this article is available as a PDF (266.4 KB).

Selected References

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

  1. Benson D. A., Boguski M., Lipman D. J., Ostell J. GenBank. Nucleic Acids Res. 1994 Sep;22(17):3441–3444. doi: 10.1093/nar/22.17.3441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chisholm D. A convenient moderate-scale procedure for obtaining DNA from bacteriophage lambda. Biotechniques. 1989 Jan;7(1):21–23. [PubMed] [Google Scholar]
  3. Chiurazzi M., Iaccarino M. Transcriptional analysis of the glnB-glnA region of Rhizobium leguminosarum biovar viciae. Mol Microbiol. 1990 Oct;4(10):1727–1735. doi: 10.1111/j.1365-2958.1990.tb00550.x. [DOI] [PubMed] [Google Scholar]
  4. Espin G., Alvarez-Morales A., Cannon F., Dixon R., Merrick M. Cloning of the glnA, ntrB and ntrC genes of Klebsiella pneumoniae and studies of their role in regulation of the nitrogen fixation (nif) gene cluster. Mol Gen Genet. 1982;186(4):518–524. doi: 10.1007/BF00337959. [DOI] [PubMed] [Google Scholar]
  5. Fitzmaurice W. P., Saari L. L., Lowery R. G., Ludden P. W., Roberts G. P. Genes coding for the reversible ADP-ribosylation system of dinitrogenase reductase from Rhodospirillum rubrum. Mol Gen Genet. 1989 Aug;218(2):340–347. doi: 10.1007/BF00331287. [DOI] [PubMed] [Google Scholar]
  6. Foster-Hartnett D., Kranz R. G. The Rhodobacter capsulatus glnB gene is regulated by NtrC at tandem rpoN-independent promoters. J Bacteriol. 1994 Aug;176(16):5171–5176. doi: 10.1128/jb.176.16.5171-5176.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fu H. A., Hartmann A., Lowery R. G., Fitzmaurice W. P., Roberts G. P., Burris R. H. Posttranslational regulatory system for nitrogenase activity in Azospirillum spp. J Bacteriol. 1989 Sep;171(9):4679–4685. doi: 10.1128/jb.171.9.4679-4685.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hartmann A., Burris R. H. Regulation of nitrogenase activity by oxygen in Azospirillum brasilense and Azospirillum lipoferum. J Bacteriol. 1987 Mar;169(3):944–948. doi: 10.1128/jb.169.3.944-948.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hartmann A., Fu H., Burris R. H. Regulation of nitrogenase activity by ammonium chloride in Azospirillum spp. J Bacteriol. 1986 Mar;165(3):864–870. doi: 10.1128/jb.165.3.864-870.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kanemoto R. H., Ludden P. W. Amino acid concentrations in Rhodospirillum rubrum during expression and switch-off of nitrogenase activity. J Bacteriol. 1987 Jul;169(7):3035–3043. doi: 10.1128/jb.169.7.3035-3043.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kanemoto R. H., Ludden P. W. Effect of ammonia, darkness, and phenazine methosulfate on whole-cell nitrogenase activity and Fe protein modification in Rhodospirillum rubrum. J Bacteriol. 1984 May;158(2):713–720. doi: 10.1128/jb.158.2.713-720.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kranz R. G., Pace V. M., Caldicott I. M. Inactivation, sequence, and lacZ fusion analysis of a regulatory locus required for repression of nitrogen fixation genes in Rhodobacter capsulatus. J Bacteriol. 1990 Jan;172(1):53–62. doi: 10.1128/jb.172.1.53-62.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Li J. D., Hu C. Z., Yoch D. C. Changes in amino acid and nucleotide pools of Rhodospirillum rubrum during switch-off of nitrogenase activity initiated by NH4+ or darkness. J Bacteriol. 1987 Jan;169(1):231–237. doi: 10.1128/jb.169.1.231-237.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Liang J. H., Nielsen G. M., Lies D. P., Burris R. H., Roberts G. P., Ludden P. W. Mutations in the draT and draG genes of Rhodospirillum rubrum result in loss of regulation of nitrogenase by reversible ADP-ribosylation. J Bacteriol. 1991 Nov;173(21):6903–6909. doi: 10.1128/jb.173.21.6903-6909.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Liang Y. Y., Arsène F., Elmerich C. Characterization of the ntrBC genes of Azospirillum brasilense Sp7: their involvement in the regulation of nitrogenase synthesis and activity. Mol Gen Genet. 1993 Aug;240(2):188–196. doi: 10.1007/BF00277056. [DOI] [PubMed] [Google Scholar]
  16. Ludden P. W., Roberts G. P. Regulation of nitrogenase activity by reversible ADP ribosylation. Curr Top Cell Regul. 1989;30:23–56. doi: 10.1016/b978-0-12-152830-0.50004-9. [DOI] [PubMed] [Google Scholar]
  17. Martin G. B., Chapman K. A., Chelm B. K. Role of the Bradyrhizobium japonicum ntrC gene product in differential regulation of the glutamine synthetase II gene (glnII). J Bacteriol. 1988 Dec;170(12):5452–5459. doi: 10.1128/jb.170.12.5452-5459.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Martin G. B., Thomashow M. F., Chelm B. K. Bradyrhizobium japonicum glnB, a putative nitrogen-regulatory gene, is regulated by NtrC at tandem promoters. J Bacteriol. 1989 Oct;171(10):5638–5645. doi: 10.1128/jb.171.10.5638-5645.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Masepohl B., Krey R., Klipp W. The draTG gene region of Rhodobacter capsulatus is required for post-translational regulation of both the molybdenum and the alternative nitrogenase. J Gen Microbiol. 1993 Nov;139(11):2667–2675. doi: 10.1099/00221287-139-11-2667. [DOI] [PubMed] [Google Scholar]
  20. Minamide L. S., Bamburg J. R. A filter paper dye-binding assay for quantitative determination of protein without interference from reducing agents or detergents. Anal Biochem. 1990 Oct;190(1):66–70. doi: 10.1016/0003-2697(90)90134-u. [DOI] [PubMed] [Google Scholar]
  21. Nordlund S., Kanemoto R. H., Murrell S. A., Ludden P. W. Properties and regulation of glutamine synthetase from Rhodospirillum rubrum. J Bacteriol. 1985 Jan;161(1):13–17. doi: 10.1128/jb.161.1.13-17.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Soliman A., Nordlund S. Purification and partial characterization of glutamine synthetase from the photosynthetic bacterium Rhodospirillum rubrum. Biochim Biophys Acta. 1989 Feb 2;994(2):138–141. doi: 10.1016/0167-4838(89)90152-0. [DOI] [PubMed] [Google Scholar]
  23. Szeto W. W., Nixon B. T., Ronson C. W., Ausubel F. M. Identification and characterization of the Rhizobium meliloti ntrC gene: R. meliloti has separate regulatory pathways for activation of nitrogen fixation genes in free-living and symbiotic cells. J Bacteriol. 1987 Apr;169(4):1423–1432. doi: 10.1128/jb.169.4.1423-1432.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Toukdarian A., Kennedy C. Regulation of nitrogen metabolism in Azotobacter vinelandii: isolation of ntr and glnA genes and construction of ntr mutants. EMBO J. 1986 Feb;5(2):399–407. doi: 10.1002/j.1460-2075.1986.tb04225.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  26. Woehle D. L., Lueddecke B. A., Ludden P. W. ATP-dependent and NAD-dependent modification of glutamine synthetase from Rhodospirillum rubrum in vitro. J Biol Chem. 1990 Aug 15;265(23):13741–13749. [PubMed] [Google Scholar]
  27. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  28. Zhang Y., Burris R. H., Ludden P. W., Roberts G. P. Comparison studies of dinitrogenase reductase ADP-ribosyl transferase/dinitrogenase reductase activating glycohydrolase regulatory systems in Rhodospirillum rubrum and Azospirillum brasilense. J Bacteriol. 1995 May;177(9):2354–2359. doi: 10.1128/jb.177.9.2354-2359.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Zhang Y., Burris R. H., Ludden P. W., Roberts G. P. Posttranslational regulation of nitrogenase activity by anaerobiosis and ammonium in Azospirillum brasilense. J Bacteriol. 1993 Nov;175(21):6781–6788. doi: 10.1128/jb.175.21.6781-6788.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zhang Y., Burris R. H., Ludden P. W., Roberts G. P. Posttranslational regulation of nitrogenase activity in Azospirillum brasilense ntrBC mutants: ammonium and anaerobic switch-off occurs through independent signal transduction pathways. J Bacteriol. 1994 Sep;176(18):5780–5787. doi: 10.1128/jb.176.18.5780-5787.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Zhang Y., Burris R. H., Roberts G. P. Cloning, sequencing, mutagenesis, and functional characterization of draT and draG genes from Azospirillum brasilense. J Bacteriol. 1992 May;174(10):3364–3369. doi: 10.1128/jb.174.10.3364-3369.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zinchenko V., Churin Y., Shestopalov V., Shestakov S. Nucleotide sequence and characterization of the Rhodobacter sphaeroides glnB and glnA genes. Microbiology. 1994 Aug;140(Pt 8):2143–2151. doi: 10.1099/13500872-140-8-2143. [DOI] [PubMed] [Google Scholar]
  33. de Bruijn F. J., Rossbach S., Schneider M., Ratet P., Messmer S., Szeto W. W., Ausubel F. M., Schell J. Rhizobium meliloti 1021 has three differentially regulated loci involved in glutamine biosynthesis, none of which is essential for symbiotic nitrogen fixation. J Bacteriol. 1989 Mar;171(3):1673–1682. doi: 10.1128/jb.171.3.1673-1682.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. de Zamaroczy M., Paquelin A., Elmerich C. Functional organization of the glnB-glnA cluster of Azospirillum brasilense. J Bacteriol. 1993 May;175(9):2507–2515. doi: 10.1128/jb.175.9.2507-2515.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES