Skip to main content
Log in

Repressor properties of the nifL gene product in Klebsiella pneumoniae

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Certain mutations in the nifL gene of the Klebsiella pneumoniae nitrogen fixation (nif) gene cluster resulted in altered nif regulaiton such that nitrogenase synthesis was no longer repressed by low levels of exogenous fixed nitrogen, by oxygen or by high temperature. Introduction of a plasmid with a nifL + allele restored fixed nitrogen and oxygen repression. We therefore conclude that the nifL product acts as a nif-specific repressor in response to these effectors.

Hence nif-specific regulation is controlled by the products of two regulatory genes, nifLA, which comprise a single operon. As previously reported (Dixon et al. 1980; Buchanan-Wollaston et al. 1981), the nifA product is necessary for transcription from other nif promoters but not from that of its own operon. We find that the nifL gene product also acts at other nif promoters but does not repress its own synthesis. Transcription of nifLA is in turn controlled by a general nitrogen-regulatory system in the cell, mediated by the products of the ntrA (glnF), ntrB (glnL) and ntrC (glnG) genes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Ausubel FM, Cannon FC (1981) Molecular genetic analysis of Klebsiella pneumoniae nitrogen fixation (nif) genes. Cold Spring Harbor Symp Quant Biol XLV:487–499

    Google Scholar 

  • Ausubel FM, Margolskee RF, Maizels NM (1977a) Mutants of Klebsiella pneumoniae in which expression of nitrogenase is independent of glutamine synthetase control. In: Newton W, Postgate JR, Rodriguez-Barrueco C (eds) Recent developments in nitrogen fixation. Academic Press, New York, p 347–356

    Google Scholar 

  • Ausubel FM, Riedel G, Cannon F, Peskin A, Margolskee R (1977b) Cloning nitrogen fixing genes from Klebsiella pneumoniae in vitro and the isolation of nif promoter mutants affecting glutamine synthetase regulation. In: Hollaender A (ed) Genetic Engineering for nitrogen fixation. Plenum Press, New York, p 111–128

    Google Scholar 

  • Brill WJ, Steiner AL, Shah VK (1974) Effect of molybdenum starvation and tungsten on the synthesis of nitrogenase components in Klebsiella pneumoniae. J Bacteriol 118:986–989

    Google Scholar 

  • Buchanan-Wollaston V, Cannon MC, Beynon J, Cannon FC (1981) The role of the nifA gene product in the transcriptional regulation of nif (nitrogen fixation) expression in Klebsiella pneumoniae. Nature 294:776–778

    Google Scholar 

  • Dixon RA, Cannon FC, Kondorosi A (1976) Construction of a P plasmid carrying nitrogen fixation genes from Klebsiella pneumoniae. Nature 260:268–271

    Google Scholar 

  • Dixon R, Eady R, Espin G, Hill S, Iaccarino M, Kahn D, Merrick M (1980) Analysis of regulation of the Klebsiella pneumoniae nitrogen fixation (nif) gene cluster with gene fusions. Nature 286:128–132

    Google Scholar 

  • Dixon R, Kennedy C, Kondorosi A, Krishnapillai V, Merrick M (1977) Complementation analysis of Klebsiella pneumoniae mutants defective in nitrogen fixation. Mol Gen Genet 157:189–198

    Google Scholar 

  • Eady RR, Issack R, Kennedy C, Postgate JR, Ratcliffe H (1978) Nitrogenase synthesis in Klebsiella pneumoniae: comparison of ammonium and oxygen regulation. J Gen Microbiol 104:277–285

    Google Scholar 

  • Eady RR, Smith BE, Cook KA, Postgate JR (1972) Nitrogenase of Klebsiella pneumoniae: purification and properties of the component proteins. Biochem J 128:655–675

    Google Scholar 

  • Elmerich C, Houmard J, Sibold L, Manheimer I, Charpin N (1978) Genetic and biochemical analysis of mutants induced by bacteriophage Mu DNA integration into Klebsiella pneumoniae nitrogen fixation genes. Mol Gen Genet 165:181–189

    Google Scholar 

  • Hennecke H, Shanmugam KT (1979) Temperature control of nitrogen fixation in Klebsiella pneumoniae. Arch Microbiol 123:259–265

    Google Scholar 

  • Hill S, Kavanagh E (1980) Roles of nifF and nifF gene products in electron transport to nitrogenase in Klebsiella pneumoniae. J Bacteriol 141:470–475

    Google Scholar 

  • Hill S, Kennedy C, Kavanagh E, Goldberg RB, Hanau R (1981) Nitrogen fixation gene (nifL) involved in oxygen regulation of nitrogenase synthesis in K. pneumoniae. Nature 290:424–426

    Google Scholar 

  • Hom SSM, Hennecke H, Shanmugam KT (1980) Regulation of nitrogenase biosynthesis in Klebsiella pneumoniae: effect of nitrate. J gen Microbiol 117:169–179

    Google Scholar 

  • Houmard J, Bogusz D, Bigault R, Elmerich C (1980) Characterisation and kinetics of the biosynthesis of some nitrogen fixation (nif) gene products in Klebsiella pneumoniae. Biochimie 62:267–275

    Google Scholar 

  • Janssen KA, Reidel GE, Ausubel FM, Cannon FC (1980) Transcriptional studies with cloned nitrogen fixation genes. In: Newton WE, Orme-Johnson WH (eds) Nitrogen fixation, vol 1. University Park Press, Baltimore, pp 85–93

    Google Scholar 

  • Kahn D, Hawkins M, Eady R (1982) Nitrogen fixation in Klebsiella pneumoniae: Nitrogenase levels and the effect of added molybdate on nitrogenase derepressed under Mo-deprivation. J gen Microbiol (in press)

  • Kennedy C (1977) Linkage map of the nitrogen fixation (nif) genes in Klebsiella pneumoniae. Mol Gen Genet 157:199–204

    Google Scholar 

  • Kennedy C, Cannon F, Cannon M, Dixon R, Hill S, Jensen J, Kumar S, McLean P, Merrick M, Robson R, Postgate J (1981) Recent advances in the genetics and regulation of nitrogen fixation. In: Gibson AH, Newton WE (eds) Current perspectives in nitrogen fixation. Australian Academy of Sciences, Canberra, pp 146–156

    Google Scholar 

  • Kennedy C, Eady RR, Kondorosi E, Klavans Rekosh D (1976) The molybdenum-iron protein of Klebsiella pneumoniae: evidence for non-identical subunits from peptide mapping. Biochem J 155:383–389

    Google Scholar 

  • MacNeil T, MacNeil D, Roberts GP, Supiano MA, Brill WJ (1978) Fine.structure mapping and complementation analysis of nif (nitrogen fixation) genes in Klebsiella pneumoniae. J Bacteriol 136:253–266

    Google Scholar 

  • MacNeil D, Brill WJ (1980) Mutations in nif genes that cause Klebsiella pneumoniae to be derepressed for nitrogenase synthesis in the presence of ammonium. J Bacteriol 144:744–751

    Google Scholar 

  • MacNeil D, Zhu J, Brill WJ (1981) Regulation of nitrogen fixation in Klebsiella pneumoniae: Isolation and characterisation of strains with nif-lac fusions. J Bacteriol 145:348–357

    Google Scholar 

  • McFarland N, McCarter L, Artz S, Kustu S (1981) Nitrogen regulatory locus ‘glnR’ of enteric bacteria is composed of cistrons ntrB and ntrC: Identification of their protein products. Proc Natl Acad Sci USA 78:2135–2139

    Google Scholar 

  • McLean P, Dixon R (1981) Requirement of nifV gene for production of wild-type nitrogenase enzyme in Klebsiella pneumoniae. Nature 292:655–656

    Google Scholar 

  • Merrick M, Filser M, Dixon R, Elmerich C, Sibold L, Houmard J (1980) Use of translocatable genetic elements to construct a fine-structure map of the Klebsiella pneumoniae nitrogen fixation (nif) gene cluster. J Gen Microbiol 117:509–520

    Google Scholar 

  • Merrick M, Filser M, Kennedy C, Dixon R (1978) Polarity of mutations induced by insertion of transposons Tn5, Tn7 and Tn10 into the nif gene cluster of Klebsiella pneumoniae. Mol Gen Genet 165:103–111

    Google Scholar 

  • Nieva-Gomez D, Roberts GP, Klevickis S, Brill WJ (1980) Electron transport to nitrogenase in Klebsiella pneumoniae. Proc Natl Acad Sci USA 77:2555–2558

    Google Scholar 

  • O'Farrell PH (1975) high resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021

    Google Scholar 

  • Pühler A, Klipp W (1981) Fine-structure analysis of the gene region for N2-fixation (nif) of Klebsiella pneumoniae. In: Boethe H, Trebst A (eds) Biology of inorganic nitrogen and sulphur. Springer-Verlag, Berlin Heidelberg New York, pp 276–286

    Google Scholar 

  • Roberts GP, Brill WJ (1980) Gene-product relationships of the nif regulon of Klebsiella pneumoniae. J Bacteriol 144:210–216

    Google Scholar 

  • Roberts GP, MacNeil T, MacNeil D, Brill WJ (1978) Regulation and characterisation of protein products coded by the nif (nitrogen fixation) genes of Klebsiella pneumoniae. J Bacteriol 136:267–279

    Google Scholar 

  • Shanmugam KT, Morandi C (1976) Amino acids as repressors of nitrogenase biosynthesis in Klebsiella pneumoniae. Biochim Biophys Acta 408:101–111

    Google Scholar 

  • Sibold L, Melck D, Elmerich C (1981) A nif mutant of Klebsiella pneumoniae fixing nitrogen in the presence of ammonia. FEMS Microbiol Lett 10:37–41

    Google Scholar 

  • St John RT, Shah VK, Brill WJ (1974) Regulation of nitrogenase synthesis by oxygen in Klebsiella pneumoniae. J Bacteriol 119:266–269

    Google Scholar 

  • Streicher SL, Shanmugan KT, Ausubel F, Morandi C, Goldberg RB (1974) Regulation of nitrogen fixation in Klebsiella pneumoniae: evidence for a role of glutamine synthetase as a regulator of nitrogenase synthesis. J Bacteriol 120:815–821

    Google Scholar 

  • Tubb RS, Postgate JR (1973) Control of nitrogenase synthesis in Klebsiella pneumoniae. J gen Microbiol 79:103–117

    Google Scholar 

  • Tubb RS (1974) Glutamine synthetase and ammonium regulation of nitrogenase synthesis in Klebsiella. Nature 251:481–485

    Google Scholar 

  • Zhu J, Brill WJ (1981) Temperature sensitivity of the regulation of nitrogenase synthesis by Klebsiella pneumoniae. J Bacteriol 145:1116–1118

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by J. Schell

Rights and permissions

Reprints and permissions

About this article

Cite this article

Merrick, M., Hill, S., Hennecke, H. et al. Repressor properties of the nifL gene product in Klebsiella pneumoniae . Molec Gen Genet 185, 75–81 (1982). https://doi.org/10.1007/BF00333793

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00333793

Keywords

Navigation