The ammonium-inactivated cyanobacterial glutamine synthetase I is reactivated in vivo by a mechanism involving proteolytic removal of its inactivating factors.
The Synechocystis sp. PCC 6803 glutamine synthetase type I (GS) activity is controlled by a process that involves protein-protein interaction with two inactivating factors (IF7 and IF17). Following addition of ammonium, the genes encoding these proteins, gifA and gifB, respectively, are derepressed, leading to the synthesis of IF7 and IF17 and consequently GS is inactivated. Upon ammonium removal, the GS activity rapidly returns to the initial level within 20 min. In this study, we analyse the mechanism underlying GS reactivation and find that this process involves IF7 and IF17 degradation. We show that the presence of ammonium as nitrogen source enhances IF17 but not IF7 stability independently of gif gene transcription. Studies with Synechocystis crude extracts under different conditions revealed that IF7 and IF17 display different stabilities in vitro. We found that IF7 is degraded in vitro by the activity of metalloproteases. Furthermore, the involvement of soluble processing metallopeptidases in IF7 degradation has also been demonstrated in vivo, by analysing Synechocystis mutant strains devoid of genes of the prp family. Finally, using a Synechocystis strain lacking GS type I, we establish the crucial role of the target protein GS for in vivo IF7 and IF17 stability.
Galmozzi
Carla V
CV
Instituto de Bioquímica Vegetal y Fotosíntesis, CSIC-Universidad de Sevilla, Américo Vespucio 49, E-41092 Sevilla, Spain.
Fernández-Avila
M Jesús
MJ
Reyes
José C
JC
Florencio
Francisco J
FJ
Muro-Pastor
M Isabel
MI
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation; D004795:Enzyme Stability; D015964:Gene Expression Regulation, Bacterial; D005974:Glutamate-Ammonia Ligase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D045726:Metalloproteases / Q000378:metabolism*; D000644:Quaternary Ammonium Compounds / Q000378:metabolism / Q000494:pharmacology*; D046939:Synechocystis / Q000201:enzymology* / Q000378:metabolism
D001426:Bacterial Proteins; C120035:IF17 protein, Synechocystis sp.; C120034:IF7 protein, Synechocystis sp.; D000644:Quaternary Ammonium Compounds; D045726:Metalloproteases; C081173:glutamine synthetase I; D005974:Glutamate-Ammonia Ligase