NtcA represses transcription of gifA and gifB, genes that encode inhibitors of glutamine synthetase type I from Synechocystis sp. PCC 6803.
Synechocystis sp. PCC 6803 glutamine synthetase type I (GS) activity is controlled by direct interaction with two inactivating factors (IF7 and IF17). IF7 and IF17 are homologous polypeptides encoded by the gifA and gifB genes respectively. We investigated the transcriptional regulation of these genes. Expression of both genes is maximum in the presence of ammonium, when GS is inactivated. Nitrogen starvation attenuates the ammonium-mediated induction of gifA and gifB as well as the ammonium-mediated inactivation of GS. Putative binding sites for the transcription factor NtcA were identified at -7.5 and -30.5 bp upstream of gifB and gifA transcription start points respectively. Synechocystis NtcA protein binding to both promoters was demonstrated by gel electrophoresis mobility shift assays. Constitutive high expression levels of both genes were found in a Synechocystis NtcA non-segregated mutant (SNC1), which showed a fourfold reduction in the ntcA expression. These experiments indicate a repressive role for NtcA on the transcription of gifA and gifB genes. Our results demonstrate that NtcA plays a central role in GS regulation in cyanobacteria, stimulating transcription of the glnA gene (GS structural gene) and suppressing transcription of the GS inactivating factor genes gifA and gifB.
García-Domínguez
M
M
Instituto de Bioquímica Vegetal y Fotosíntesis, Centro de Investigaciones Científicas Isla de la Cartuja, C/. Américo Vespucio s/n, 41092 Sevilla, Spain.
Reyes
J C
JC
Florencio
F J
FJ
D001426:Bacterial Proteins; D001483:Base Sequence; D001665:Binding Sites; D000458:Cyanobacteria / Q000201:enzymology* / Q000235:genetics; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000378:metabolism / Q000502:physiology*; D005974:Glutamate-Ammonia Ligase / Q000235:genetics*; D007527:Isoenzymes / Q000235:genetics*; D016254:Mutagenesis, Insertional; D009584:Nitrogen / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000378:metabolism / Q000502:physiology*; D014157:Transcription Factors / Q000378:metabolism / Q000502:physiology*; D014158:Transcription, Genetic / Q000502:physiology*
D001426:Bacterial Proteins; D017931:DNA Primers; D004268:DNA-Binding Proteins; D007527:Isoenzymes; D012097:Repressor Proteins; D014157:Transcription Factors; C075617:ntcA protein, Synechococcus; D005974:Glutamate-Ammonia Ligase; D009584:Nitrogen