Motility Functions as an Essential Defense in the Bacterial Persister Lifecycle

Abstract Bacterial persisters are phenotypically antibiotic-tolerant variants driving recurrent infections. While motility is a key trait for bacterial survival and virulence, its function throughout the persister lifecycle remains poorly understood. Here, we unveil a stage-specific trajectory for motility that underlies sophisticated survival strategies in rifampin-induced Escherichia coli persistence. During persister formation, intact motility functions as an immediate behavioral defense that decreases intracellular bactericidal antibiotic concentrations to ensure survival. This initial behavioral defense provides critical time for transcriptional reprogramming in persisters, which, by downregulating the major outer membrane porins (OmpC & OmpF) that facilitate antibiotic influx, supplants the preceding motility-based strategy. Subsequently, in structurally fortified persisters, motility becomes dispensable and gradually diminishes– a process independent of cell bioenergetic profile–yet re-emerges as essential for timely resuscitation upon antibiotic removal. Collectively, these findings establish the significance of motility in bacterial survival, redefine persister defense against antibiotics as a transition from behavioral to architectural strategies, and identify motility as a potential adjuvant therapeutic target. Graphical Abstract The dynamic roles and underlying mechanisms of bacterial motility throughout the rifampin-induced persister lifecycle: motility initially serves as a behavioral defense mechanism essential for persister formation, then becoming dispensable in the persister state which implements a structural defense via membrane porin downregulation, and ultimately re-emerges as a critical requirement for timely resuscitation.