Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1978 Oct;136(1):267–279. doi: 10.1128/jb.136.1.267-279.1978

Regulation and characterization of protein products coded by the nif (nitrogen fixation) genes of Klebsiella pneumoniae.

G P Roberts, T MacNeil, D MacNeil, W J Brill
PMCID: PMC218657  PMID: 361694

Abstract

Two hundred and thirty-five Nif- strains of Klebsiella pneumoniae were characterized by two-dimensional polyacrylamide gel electrophoresis. Forty-two of these strains were tested further by in vitro acetylene reduction assays. By these techniques, nine nif-coded polypeptides were identified, and eight of these were assigned to specific nif genes. Nitrogenase component I required nifK and nifD, which coded for the beta and alpha subunits, and nifB, -E, and -N were required for the iron-molybdenum cofactor, which is a part of the active site of nitrogenase. nifH coded for the structural protein of component II, and nifM and nifS products seemed to be necessary for the synthesis of an active component II. There were two genes, nifF and nifJ, that were required for N2 fixation in vivo but not for N2 fixation in vitro. There were at least two cases (nifE and nifN, nifK and nifD) of two proteins that seemed to require each other for stability in vivo. Regulation of N2 fixation is apparently complex, and this is reflected by the assignment of regulatory functions to the gene products of nifA, nifL, nifK, nifD, nifH, and NIFJ.

Full text

PDF
274

Images in this article

269Image
on p.269
273Image
on p.273
274Image
on p.274
274Image
on p.274
275Image
on p.275
276Image
on p.276
277Image
on p.277
277Image
on p.277

Selected References

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

  1. Ausubel F., Riedel G., Cannon F., Peskin A., Margolskee R. Cloning nitrogen fixing genes from Klebsiella pneumoniae in vitro and the isolation of NIF promoter mutants affecting glutamine synthetase regulation. Basic Life Sci. 1977;9:111–128. doi: 10.1007/978-1-4684-0880-5_10. [DOI] [PubMed] [Google Scholar]
  2. Brill W. J., Steiner A. L., Shah V. K. Effect of molybdenum starvation and tungsten on the synthesis of nitrogenase components in Klebsiella pneumonia. J Bacteriol. 1974 Jun;118(3):986–989. doi: 10.1128/jb.118.3.986-989.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Collmer A., Lamborg M. Arrangement and regulation of the nitrogen fixation genes in Klebsiella pneumoniae studied by depression kinetics. J Bacteriol. 1976 May;126(2):806–813. doi: 10.1128/jb.126.2.806-813.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dixon R., Kennedy C., Kondorosi A., Krishnapillai V., Merrick M. Complementation analysis of Klebsiella pneumoniae mutants defective in nitrogen fixation. Mol Gen Genet. 1977 Nov 29;157(2):189–198. doi: 10.1007/BF00267397. [DOI] [PubMed] [Google Scholar]
  5. Eady R. R., Smith B. E., Cook K. A., Postgate J. R. Nitrogenase of Klebsiella pneumoniae. Purification and properties of the component proteins. Biochem J. 1972 Jul;128(3):655–675. doi: 10.1042/bj1280655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kennedy C., Eady R. R., Kondorosi E., Rekosh D. K. The molybdenum--iron protein of Klebsiella pneumoniae nitrogenase. Evidence for non-identical subunits from peptide 'mapping'. Biochem J. 1976 May 1;155(2):383–389. doi: 10.1042/bj1550383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kennedy C. Linkage map of the nitrogen fixation (nif) genes in Klebsiella pneumoniae. Mol Gen Genet. 1977 Nov 29;157(2):199–204. doi: 10.1007/BF00267398. [DOI] [PubMed] [Google Scholar]
  8. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  9. MacNeil T., Brill W. J., Howe M. M. Bacteriophage mu-induced deletions in a plasmid containing the nif (N2 fixation) genes of Klebsiella pneumoniae. J Bacteriol. 1978 Jun;134(3):821–829. doi: 10.1128/jb.134.3.821-829.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. MacNeil T., MacNeil D., Roberts G. P., Supiano M. A., Brill W. J. Fine-structure mapping and complementation analysis of nif (nitrogen fixation) genes in Klebsiella pneumoniae. J Bacteriol. 1978 Oct;136(1):253–266. doi: 10.1128/jb.136.1.253-266.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  12. Shah V. K., Brill W. J. Isolation of an iron-molybdenum cofactor from nitrogenase. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3249–3253. doi: 10.1073/pnas.74.8.3249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Shah V. K., Brill W. J. Nitrogenase. IV. Simple method of purification to homogeneity of nitrogenase components from Azotobacter vinelandii. Biochim Biophys Acta. 1973 May 30;305(2):445–454. doi: 10.1016/0005-2728(73)90190-4. [DOI] [PubMed] [Google Scholar]
  14. Shah V. K., Davis I. C., Gordon J. K., Orme-Johnson W. H., Brill W. J. Nitrogenase. 3. Nitrogenaseless mutants of Azotobacter vinelandii: activities, cross-reactions and EPR spectra. Biochim Biophys Acta. 1973 Jan 18;292(1):246–255. doi: 10.1016/0005-2728(73)90269-7. [DOI] [PubMed] [Google Scholar]
  15. St John R. T., Johnston H. M., Seidman C., Garfinkel D., Gordon J. K., Shah V. K., Brill W. J. Biochemistry and genetics of Klebsiella pneumoniae mutant strains unable to fix N2. J Bacteriol. 1975 Mar;121(3):759–765. doi: 10.1128/jb.121.3.759-765.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. St John R. T., Shah V. K., Brill W. J. Regulation of nitrogenase synthesis by oxygen in Klebsiella pneumoniae. J Bacteriol. 1974 Jul;119(1):266–269. doi: 10.1128/jb.119.1.266-269.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Streicher S. L., Shanmugam K. T., Ausubel F., Morandi C., Goldberg R. B. Regulation of nitrogen fixation in Klebsiella pneumoniae: evidence for a role of glutamine synthetase as a regulator of nitrogenase synthesis. J Bacteriol. 1974 Nov;120(2):815–821. doi: 10.1128/jb.120.2.815-821.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tubb R. S. Glutamine synthetase and ammonium regulation of nitrogenase synthesis in Klebsiella. Nature. 1974 Oct 11;251(5475):481–485. doi: 10.1038/251481a0. [DOI] [PubMed] [Google Scholar]
  19. Tubb R. S., Postgate J. R. Control of nitrogenase synthesis in Klebsiella pneumoniae. J Gen Microbiol. 1973 Nov;79(1):103–117. doi: 10.1099/00221287-79-1-103. [DOI] [PubMed] [Google Scholar]
  20. Yoch D. C., Pengra R. M. Effect of amino acids on the nitrogenase system of Klebsiella pneumoniae. J Bacteriol. 1966 Sep;92(3):618–622. doi: 10.1128/jb.92.3.618-622.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES