An intracellular phosphorus-starvation signal activates the PhoB/PhoR two-component system in Salmonella enterica

Bacteria acquire P primarily as inorganic orthophosphate (Pi, PO₄^3-). Once internalized, Pi is rapidly assimilated into biomass during the synthesis of ATP. Because Pi is essential, but excessive ATP is toxic, the acquisition of environmental Pi is tightly regulated. In the bacterium Salmonella enterica (Salmonella), growth in Pi-limiting environments activates the membrane sensor histidine kinase PhoR, leading to the phosphorylation of its cognate transcriptional regulator PhoB and subsequent transcription of genes involved in adaptations to low Pi. Pi limitation promotes PhoR kinase activity by altering the conformation of a membrane signaling complex comprised of PhoR, the multicomponent Pi transporter system PstSACB and the regulatory protein PhoU. However, the identity of the Pi-starvation signal and how it controls PhoR activity remain unknown. Here, we identify conditions where the PhoB and PhoR signal transduction proteins can be maintained in an inactive state when Salmonella is grown in media lacking Pi. Our results demonstrate that PhoB/PhoR is activated by an intracellular P-insufficiency signal.IMPORTANCEIn enteric bacteria, the transcriptional response to phosphorus (P) starvation is controlled by a specialized signal transduction system comprised of a membrane-bound, multicomponent signal sensor, and a cytoplasmic transcriptional factor. Whereas this system has been primarily studied in the context of phosphate (Pi) starvation, it is currently unknown how this stress initiates signal transduction. In the current study, we establish that this signaling system is regulated by a cytoplasmic signal arising from insufficient P. We demonstrate that rather than responding to extracellular conditions, cells couple the activation of their P starvation response to the availability of cytoplasmic P. This regulatory logic may enable cells to prevent toxicity resulting from excessive Pi acquisition and hinder the onset of a P starvation response when their metabolic demands are being met through the consumption of P sources other than Pi.