Abstract
The unicellular cyanobacterium Synechococcus PCC6301 lacks a hybridisable homologue of the strongly conserved gdhA gene of E. coli that encodes NADP-specific glutamate dehydrogenase. This is consistent with the failure to find this enzyme in extracts of the cyanobacterium. The E. coli gdhA gene was transferred to Synechococcus PCC6301 by transformation with an integrative vector. High levels of glutamate dehydrogenase activity, similar to those found in ammonium grown E. coli cells, were found in these transformants. These transformed cyanobacteria displayed an ammonium tolerant phenotype, consistent with the action of their acquired glutamate dehydrogenase activity as an ammonium detoxification mechanism. Minor differences in colony size and in growth at low light intensity were also observed.
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References
Beevers L, Hageman RH: Nitrate and nitrite reduction. In: Miflin BJ (ed.) The Biochemistry of Plants, Volume 5, pp. 115–168. Academic Press, New York (1980).
Bottomly PJ, vanBalen C: Characterization of heterotrophic growth in the blue-green alga Nostoc sp. strain MAC. J Gen Microbiol 107: 309–318 (1978).
Bravo A, Becerril B, Mora J: Introduction of the Escherichia coli gdhA gene into Rhizobium phaseoli: effect on nitrogen fixation. J Bact 170: 985–988 (1988).
Givan CV: Metabolic detoxification of ammonia in higher plants. Phytochem 18: 375–382 (1979).
Hanahan D: Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166: 557–580 (1983).
Haselkorn R: Organisation of the genes for nitrogen fixation in photosynthetic bacteria and cyanobacteria. Ann Rev Microbiol 40: 525–547 (1986).
Hoare DS, Hoare SL, Moore RB: The photoassimilation of organic compounds by autotrophic blue-green algae. J Gen Microbiol 49: 351–370 (1967).
Ish-Horowicz D, Burke JF: Rapid and efficient cosmid vector cloning. Nucl Acids Res 9: 2989–2998 (1981).
Janson CA, Kayne PS, Almassy RJ, Grunstein M, Eisenberg D: Sequence of glutamine synthetase from Salmonella typhimurium and implications for the protein structure. Gene 46: 297–300 (1986).
Krogmann DW, Jagendorf AT, Avron M: Uncouplers of spinach chloroplast photosynthetic phosphorylation. Plant Physiol 34: 272–277 (1959).
Lightfoot DA, Walters DE, Wootton JC: Transformation of the cyanobacterium Synechococcus PCC6301 using cloned DNA. J Gen Microbiol 134: 1509–1514 (1988).
Maniatis T, Fritsch E, Sambrook J: Molecular Cloning: A Laboratory Manual, 1st edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York (1982).
Mattaj IW, McPherson MJ, Wootton JC: Localization of a strongly conserved section of coding sequence in glutamate dehydrogenase genes. FEBS Lett 147: 21–25 (1982).
Mazur BJ, Rice D, Haselkorn R: Identification of blue-green algal nitrogen fixation genes by using heterologous DNA hybridization probes. Proc Natl Acad Sci USA 77: 186–190 (1980).
McPherson MJ, Wootton JC: Complete nucleotide sequence of the Escherichia coli gdhA gene. Nucl Acids Res 11: 5257–5266 (1983).
Meeks JC, Wolk CP, Lockau W, Schilling N, Shaffer W, Chen WS: Pathways of assimilation of [13N]N2 and [13N]NH4 + by cyanobacteria with and without heterocysts. J Bact 134: 125–130 (1976).
Meister A: Catalytic mechanism of glutamine synthetase: overview of glutamine metabolism. In: Mora J, Palacios R (eds) Glutamine: Metabolism, Enzymes and Regulation, pp. 1–40. Academic Press, New York (1980).
Miflin BJ, Lea PJ: Ammonia assimilation. In: Miflin BJ (ed.) The Biochemistry of Plants, Volume 5, pp. 169–202. Academic Press, New York (1980).
Mountain A, McPherson MJ, Baron AJ, Wootton JC: The Klebsiella aerogenes glutamate dehydrogenase gene: cloning, high level expression and hybrid enzyme formation in Escherichia coli. Mol Gen Genet 199: 141–145 (1985).
Mulligan B, Schultes N, Chen L, Bogorad L: Nucleotide sequence of a multi-copy gene for the B-protein of photosystem II of a cyanobacterium. Proc Nat Acad Sci USA 81: 2693–2697 (1984).
Neilsonn AH, Doudoroff M: Ammonia assimilation in blue-green algae. Archiv Microbiol 89: 15–22 (1973).
Pearce J, Leach CK, Carr NG: The incomplete tricarboxylic acid cycle in Anabaena variabilis. J Gen Microbiol 55: 371–378 (1969).
Platt SG, Anthon GE: Ammonia accumulation and inhibition of photosynthesis in methionine sulphoxamine treated spinach. Plant Physiol 67: 509–513 (1981).
Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY: Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 111: 1–61 (1979).
Rocha M, Vazquez M, Garciarrubio A, Covarrubias AA. Nucletide sequences of the glnA-glnL intercistronic region of Escherichia coli. Gene 37: 91–99 (1985).
Sinden SL, Durbin RD: Glutamine synthetase inhibition: possible mode of action of wildfire toxin from Pseudomonas tabaci. Nature 219: 379–380 (1968).
Singh HN, Ladha JK, Kumar HD: Genetic control of heterocyst formation in the blue-green algae Nostoc muscorum and Nostoc linkea. Archiv Microbiol 144: 155–159 (1987).
Singh HN, Sonie KC, Singh HN: Nitrate regulation of heterocyst differentiation and nitrogen fixation in a chlorate resistant mutant of the blue-green alga Nostoc muscorum. Mutation Res 42: 447–452 (1977).
Smith FA, Raven JA: Intracellular pH and its regulation. Ann Rev Plant Physiol 30: 289–311 (1979).
Stanier RY, Adelberg EA, Ingraham JL: General Microbiology, 4th edn. McMillan Press, London (1977).
Stewart WDP: Some aspects of structure and function in N2-fixing cyanobacteria. Ann Rev Microbiol 34: 497–536 (1980).
Stewart GR, Mann AF, Fentem PA: Enzymes of glutamate formation: glutamate dehydrogenase, glutamine synthetase and glutamate synthase. In: Miflin BJ (ed.) The Biochemistry of Plants, Volume 5, pp. 169–202. Academic Press, New York (1980).
Tumer NE, Robinson SJ, Haselkorn R: Different promoters for the Anabaena glutamine synthetase gene during growth using molecular or fixed nitrogen. Nature 306: 337–343 (1983).
Windass JD, Worsey MJ, Pioli EM, Pioli D, Barth PT, Atherton KT, Byrom D, Powell K, Senior PJ: Inproved conversion of methanol to single-cell protein by Methylophilus methylotrophus. Nature 287: 396–401 (1981).
Wootton JC: Reassessment of ammonium ion affinities of NADP-specific glutamate dehydrogenase: activation of the Neurospora crassa enzyme by ammonium and rubidium ions. Biochem J 209: 527–531 (1983).
Wootton JC, McPherson MJ: Genes of nitrate and ammonium assimilation. Ann Proc Phytochem Soc Europe 23: 89–114 (1984).
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Lightfoot, D.A., Baron, A.J. & Wootton, J.C. Expression of the Escherichia coli glutamate dehydrogenase gene in the cyanobacterium Synechococcus PCC6301 causes ammonium tolerance. Plant Mol Biol 11, 335–344 (1988). https://doi.org/10.1007/BF00027390
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DOI: https://doi.org/10.1007/BF00027390