Phosphate limitation triggers Fe³⁺-dependent polymyxin resistance in Enterobacteriaceae

Phosphate (Pi) limitation is a pervasive stressor in bacterial habitats that often promotes increased antibiotic resistance. However, the ecophysiological drivers and regulatory mechanisms remain largely unresolved, impeding targeted interventions. Here, we identify a conserved Pi–Mg–Fe–PmrAB signalling pathway in Enterobacteriaceae that mediates Pi starvation-induced resistance to polymyxin B, a last-resort antibiotic. We demonstrate that Pi depletion disrupts Mg²⁺ homeostasis, leading to Fe³⁺ mobilization to cell membrane, which activates PmrAB and the downstream arnABCDTEF operon. This system catalyzes L-Ara4N modification of lipid A, a universal component of Gram-negative outer membranes, conferring polymyxin resistance. Unlike phosphorus-free lipid strategies in P. aeruginosa , the Mg–Fe–PmrAB pathway is druggable. We show that Mg²⁺ supplementation or Fe³⁺ chelation effectively reverses resistance. Phylogenetic analyses reveal species-specific differences in PmrB sensing capabilities, highlighting the importance of understanding regulatory diversity. Our findings uncover a conserved mechanism of stress-induced resistance, offering new avenues for resistance management.