Fine-tuning of amino sugar homeostasis by EIIA Ntr in Salmonella Typhimurium

Abstract

The nitrogen-metabolic phosphotransferase system, PTSNtr, consists of the enzymes INtr, NPr and IIANtr that are encoded by ptsP, ptsO, and ptsN, respectively. Due to the proximity of ptsO and ptsN to rpoN, the PTSNtr system has been postulated to be closely related with nitrogen metabolism. To define the correlation between PTSNtr and nitrogen metabolism, we performed ligand fishing with EIIANtr as a bait and revealed that D-glucosamine-6-phosphate synthase (GlmS) directly interacted with EIIANtr. GlmS, which converts D-fructose-6-phosphate (Fru6P) into D-glucosamine-6-phosphate (GlcN6P), is a key enzyme producing amino sugars through glutamine hydrolysis. Amino sugar is an essential structural building block for bacterial peptidoglycan and LPS. We further verified that EIIANtr inhibited GlmS activity by direct interaction in a phosphorylation-state-dependent manner. EIIANtr was dephosphorylated in response to excessive nitrogen sources and was rapidly degraded by Lon protease upon amino sugar depletion. The regulation of GlmS activity by EIIANtr and the modulation of glmS translation by RapZ suggest that the genes comprising the rpoN operon play a key role in maintaining amino sugar homeostasis in response to nitrogen availability and the amino sugar concentration in the bacterial cytoplasm.