Summary
A series of mutants defective in nitrogen fixation (nif) were isolated in Klebsiella pneunoniae strain M5a1. The nif mutations were either located on plasmid pRD1 or on the K. pneumoniae chromosome. A total of 37 plasmid mutants and 28 chromosomal mutants were employed in complementation tests using the acetylene reduction technique. Most mutants could be assigned to one of seven nif cistrons: nifA, nifB, nifD, nifE, nifF, nifH, and nifK.
Complementation analysis of two nif deletion mutants confirmed transductional evidence that these strains carry nifB-A-F deletions. One deletion mutant had, in contrast to previous transductional analysis, a functional nifK cistron and presumably is deleted for nifB-A-F-E.
Examination of the biochemical phenotype of several mutants suggests that the nifA product has a regulatory function, and nifK, nifD and nifH are most probably the structural genes for nitrogenase.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bachhuber, M., Brill, W.J., Howe, M.M.: Use of bacteriophage Mu to isolate deletions in the his-nif region of Klebsiella pneumoniae. J. Bact. 128, 749–753 (1976)
Bachman B.J., Low, K.B., Taylor, A.L.: Recalibrated linkage map of Escherichia coli K12. Bact. Rev. 40, 116–167 (1976)
Cannon, F.C., Dixon, R.A., Postgate, J.R.: Derivation and properties of F-prime factors in Escherichia coli carrying nitrogen fixation genes from Klebsiella pneumoniae. J. gen. Microbiol. 93, 111–125 (1976)
Cannon, F.C., Dixon, R.A., Postgate, J.R., Primrose, S.B.: Chromosomal integration of Klebsiella nitrogen fixation genes in Escherichia coli. J. gen. Microbiol 80, 227–239 (1974)
Cannon, F.C., Reidel, G.R., Ausubel F.M.: A recombinant plasmid which carries part of the nitrogen fixation (nif) gene cluster of K. pneumoniae. Proc. nat. Acad. Sci. (Wash.) 74, 2963–2967 (1977)
Chang, G.W., Straus, D., Ames, B.N.: Enriched selection of dominant mutations: histidine operator mutations. J. Bact. 107, 578–579 (1971)
Dixon, R.A., Cannon, F.C., Kondorosi, A.: Construction of a P plasmid carrying nitrogen fixation genes from Klebsiella pneumoniae. Nature (Lond.) 260, 268–271 (1976)
Dixon, R.A., Postgate, J.R.: Genetic transfer of nitrogen fixation from Klebsiella pneumoniae to Escherichia coli. Nature (Lond.) 237, 102–103 (1972)
Eady, R.R., Issack, R., Kennedy, C., Postgate, J.R., Ratcliffe, H.D.: Nitrogenase synthesis in Klebsiella pneumoniae: Comparison of regulation by NH +4 and oxygen. J. gen Microbiol. (in press) (1977)
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. 128, 655–657 (1972)
Kennedy, C.: Linkage map of nitrogen fixation (nif) genes in Klebsiella pneumonae. Molec. gen. Genet. 157, 199–204 (1977)
Kennedy, C., Eady, R.R., Kondorosi, E., Klavans-Rekosh, D.: The molybdenum-iron protein of Klebsiella pneumoniae: evidence for non-identical sub-units from peptide mapping. Biochem. J. 155, 383–389 (1976)
Kennedy, C., Postgate, J.R.: Expression of Klebsiella pneumoniae nitrogen fixation genes in nitrate reductase mutants of Escherichia coli. J. gen. Microbiol. 98, 551–557 (1977)
Moses, R.G.: Replicative DNA synthesis in system diffusible for macromolecules. J. biol. Chem. 247, 6031–6038 (1972)
Nagatani, H.H., Shah, V.K., Brill, W.J.: Activation of inactive nitrogenase by acid-treated component I. J. Bact. 120, 697–701 (1974)
Novick, R.P., Clowes, R.C., Cohen, S.N., Curtiss III, R., Datta, N., Falkow, S.: Uniform nomenclature for bacterial plasmids: a proposal. Bact. Rev. 40, 168–189 (1976)
Oeschger, M.P., Berlyn, M.K.B.: A simple procedure for localised mutagenesis using nitrosoguanidine. Molec. gen. Genet. 134, 77–83 (1974)
Shannugan, K.T., Loo, A.S., Valentine, R.S.: Deletion mutants of nitrogen fixation in Klebsiella pneumoniae: mapping of a cluster of nif genes essential for nitrogenase activity. Biochim. biophys. Acta (Amst.) 338, 545–553 (1974)
Smith, B.E., Lang, G.: Mossbauer spectroscopy of the nitrogenase proteins from Klebsiella pneumoniae. Structural assignments and mechanistic conclusions. Biochem. J. 137, 169–180 (1974)
Streicher, S., Gurney, E.G., Valentine, R.C.: The nitrogen fixation genes. Nature (Lond.) 239, 495–499 (1972)
Streicher, S.L., Shanmugan, 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. Bact. 120, 815–821 (1974)
St. John, R.T., Johnston, M.H., 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. Bact. 121, 759–765 (1975)
Tubb, R.S., Postgate, J.R.: Control of nitrogenase synthesis in Klebsiella pneumoniae. J. gen. Microbiol. 79, 103–117 (1973)
Author information
Authors and Affiliations
Additional information
Communicated by B.A. Bridges
Rights and permissions
About this article
Cite this article
Dixon, R., Kennedy, C., Kondorosi, A. et al. Complementation analysis of Klebsiella pneumoniae mutants defective in nitrogen fixation. Molec. Gen. Genet. 157, 189–198 (1977). https://doi.org/10.1007/BF00267397
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00267397