2-Oxoglutarate increases the binding affinity of the NtcA (nitrogen control) transcription factor for the Synechococcus glnA promoter

Abstract

The cyanobacterial NtcA global nitrogen regulator belongs to the catabolite activator protein (CAP) family and activates transcription of nitrogen assimilation genes in response to nitrogen step-down. The binding affinity of NtcA towards a DNA fragment carrying the promoter of the glnA gene from Synechococcus sp. PCC 7942, analyzed in vitro by band-shift assay, was increased five-fold by 2-oxoglutarate in the presence of Mg(2+) ions. The 2-oxoglutarate effect peaked at about 0.6 mM, a rather physiological concentration for this compound under nitrogen-limiting conditions, and could be partially reproduced by 3-oxoglutarate but not by oxaloacetate or glutamate. These results suggest 2-oxoglutarate as a signal of the C to N balance of the cells to regulate NtcA activity and provide a new example of regulation in the versatile CAP family of proteins.