Summary
A number of mutants have been isolated which affect regulation of the nitrogen fixation (nif) gene cluster in Klebsiella pneumoniae and all of which are linked to glnA, the structural gene for glutamine synthetase (G.S.). These mutants were classified on the basis of their G.S. and nitrogenase activities in conditions of nitrogen limitation and excess. The plasmid R68.45 was then used to generate a number of R-primes carrying the glnA region of the K. pneumoniae chromosome. One of these R-primes (pGE10) was subsequently used in complementation analysis and by isolation of transposon-induced insertion mutations in pGE10 we have demonstrated the existence of a gene, glnG, closely linked to glnA. Mutations in glnG have a similar phenotype to glnG mutants described in Escherichia coli (Pahel and Tyler 1979) and Salmonella typhimurium (Kustu et al. 1979) in that they substantially reduce G.S. activity but are not glutamine auxotrophs. GlnG mutants have very low nitrogenase activity indicating that the glnG product may be involved in regulation of the nif gene cluster in K. pneumoniae.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ausubel FM, Bird SC, Durbin KJ, Janssen KA, Margolskee RF, Peskin AP (1979) Glutamine synthetase mutations which affect expression of nitrogen fixation genes in Klebsiella pneumoniae. J Bacteriol 140:597–606
Ausubel FM, Riedel G, Cannon F, Peskin A, Margolskee R (1977) 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
Bachmann BJ (1972) Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev 36:525–557
Barth PT, Datta N, Hedges RW, Grinter NJ (1976) Transposition of a deoxyribonucleic acid sequence encoding trimethoprim and streptomycin resistance from R483 to other replicons. J Bacteriol 125:800–810
Bender RA, Janssen KA, Resnick AD, Blumenberg M, Foor F, Magasanik B (1977) Biochemical parameters of glutamine synthetase from Klebsiella aerogenes. J Bacteriol 129:1001–1009
Brewin NJ, Beringer JE, Johnston AWB (1980) Plasmid-mediated transfer of host-range specificity between two strains of Rhizobium leguminosarum. J Gen Microbiol 120:413–420
Dixon R, Kennedy C, Kondorosi A, Krishnapillai V, Merrick M (1977) Complementation analysis of of Klebsiella pneumoniae mutants defective in nitrogen fixation. Mol Gen Genet 157:189–198
Foster TJ, Willetts NS (1977) Characterisation of transfer-deficient mutants of the R100-1 Tcs plasmid pDU202, caused by insertion of Tn10. Mol Gen Genet 156:107–114
Goldberg RB, Hanau R (1979) Relation between the adenylylation state of glutamine synthetase and the expression of other genes involved in nitrogen metabolism. J Bacteriol 137:1282–1289
Holloway BW (1979) Plasmids that mobilise bacterial chromosome. Plasmid 2:1–19
Johnston AWB, Setchell SM, Beringer JE (1978) Interspecific crosses between Rhizobium leguminosarum and R. meliloti: Formation of haploid recombinants and of R-primes. J Gen Microbiol 104:209–218
Kennedy C (1977) Linkage map of the nitrogen fixation (nif) genes in Klebsiella pneumoniae. Mol Gen Genet 157:199–204
Kustu S, Burton D, Garcia E, McCarter L, McFarland N (1979) Nitrogen control in Salmonella: Regulation by the glnR and glnF gene products. Proc Natl Acad Sci USA 76:4576–4580
Leonardo JM, Goldberg RB (1980) Regulation of nitrogen metabolism in glutamine auxotrophs of Klebsiella pneumoniae. J Bacteriol 142:99–110
Lowry OH, Rosenberg NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
McNeil 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
Magasanik B (1977) Regulation of bacterial nitrogen assimilation by glutamine synthetase. Trends in Biochem Sci 2:9–12
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
Pahel G, Tyler B (1979). A new glnA-linked regulatory gene for glutamine synthetase in Escherichia coli. Proc Natl Acad Sci USA 76:4544–4548
Shanmugam KT, Chan I, Morandi C (1975). Regulation of nitrogen fixation. Nitrogenase-derepressed mutants of Klebsiella pneumoniae. Biochim Biophys Acta 565:365–378
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
Streicher SL, Bender RA, Magasanik B (1975) Genetic control of glutamine synthetase in Klebsiella aerogenes. J Bacteriol 121:320–331
Streicher SL, Shanmugam 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
Tubb RS (1978) Glutamine synthetase and ammonium regulation of nitrogenase synthesis in Klebsiella. Nature 251:481–485
Tyler B (1978) Regulation of the assimilation of nitrogen compounds. Annu Rev Biochem 47:1127–1162
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
Espin, G., Alvarez-Morales, A. & Merrick, M. Complementation analysis of glnA-linked mutations which affect nitrogen fixation in Klebsiella pneumoniae . Molec. Gen. Genet. 184, 213–217 (1981). https://doi.org/10.1007/BF00272907
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00272907