A mutant lacking the glutamine synthetase gene (glnA) is impaired in the regulation of the nitrate assimilation system in the cyanobacterium Synechocystis sp. strain PCC 6803.
The existence in the unicellular cyanobacterium Synechocystis sp. strain PCC 6803 of two genes (glnA and glnN) coding for glutamine synthetase (GS) has been recently reported (J.C. Reyes and F.J. Florencio, J. Bacteriol. 176:1260-1267, 1994). In the current work, the regulation of the nitrate assimilation system was studied with a glnA-disrupted Synechocystis mutant (strain SJCR3) in which the only GS activity is that corresponding to the glnN product. This mutant was unable to grow in ammonium-containing medium because of its very low levels of GS activity. In the SJCR3 strain, nitrate and nitrite reductases were not repressed by ammonium, and short-term ammonium-promoted inhibition of nitrate uptake was impaired. In Synechocystis sp. strain PCC 6803, nitrate seems to act as a true inducer of its assimilation system, in a way similar to that proposed for the dinitrogen-fixing cyanobacteria. A spontaneous derivative strain from SJCR3 (SJCR3.1), was able to grow in ammonium-containing medium and exhibited a fourfold-higher level of GS activity than but the same amount of glnN transcript as its parental strain (SJCR3). Taken together, these finding suggest that SJCR3.1 is a mutant affected in the posttranscriptional regulation of the GS encoded by glnN. This strain recovered regulation by ammonium of nitrate assimilation. SJCR3 cells were completely depleted of intracellular glutamine shortly after addition of ammonium to cells growing with nitrate, while SJCR3.1 cells maintained glutamine levels similar to that reached in the wild-type Synechocystis sp. strain PCC 6803. Our results indicate that metabolic signals that control the nitrate assimilation system in Synechocystis sp. strain PCC 6803 require ammonium metabolism through GS.
Reyes
J C
JC
Departamento de Bioquímica Vegetal, Universidad de Sevilla-CSIC, Spain.
Florencio
F J
FJ
D000596:Amino Acids / Q000032:analysis; D001692:Biological Transport; D000458:Cyanobacteria / Q000737:chemistry / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005974:Glutamate-Ammonia Ligase / Q000235:genetics*; D009154:Mutation; D050901:Nitrate Reductase; D009565:Nitrate Reductases / Q000378:metabolism; D009566:Nitrates / Q000378:metabolism*; D009572:Nitrite Reductases / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D000644:Quaternary Ammonium Compounds / Q000378:metabolism
D000596:Amino Acids; D009566:Nitrates; D009573:Nitrites; D000644:Quaternary Ammonium Compounds; D009565:Nitrate Reductases; D009572:Nitrite Reductases; D050901:Nitrate Reductase; C081173:glutamine synthetase I; D005974:Glutamate-Ammonia Ligase