The Signal Transduction Protein P_II Controls Ammonium, Nitrate and Urea Uptake in Cyanobacteria

P_II signal transduction proteins are widely spread among all domains of life where they regulate a multitude of carbon and nitrogen metabolism related processes. Non-diazotrophic cyanobacteria can utilize a high variety of organic and inorganic nitrogen sources. In recent years, several physiological studies indicated an involvement of the cyanobacterial P_II protein in regulation of ammonium, nitrate/nitrite, and cyanate uptake. However, direct interaction of P_II has not been demonstrated so far. In this study, we used biochemical, molecular genetic and physiological approaches to demonstrate that P_II regulates all relevant nitrogen uptake systems in Synechocystis sp. strain PCC 6803: P_II controls ammonium uptake by interacting with the Amt1 ammonium permease, probably similar to the known regulation of E. coli ammonium permease AmtB by the P_II homolog GlnK. We could further clarify that P_II mediates the ammonium- and dark-induced inhibition of nitrate uptake by interacting with the NrtC and NrtD subunits of the nitrate/nitrite transporter NrtABCD. We further identified the ABC-type urea transporter UrtABCDE as novel P_II target. P_II interacts with the UrtE subunit without involving the standard interaction surface of P_II interactions. The deregulation of urea uptake in a P_II deletion mutant causes ammonium excretion when urea is provided as nitrogen source. Furthermore, the urea hydrolyzing urease enzyme complex appears to be coupled to urea uptake. Overall, this study underlines the great importance of the P_II signal transduction protein in the regulation of nitrogen utilization in cyanobacteria.