MotY modulates proton-driven flagellar motor output in Pseudomonas aeruginosa

MotY homologs are present in a variety of monotrichous bacterial strains and are thought to form an additional structural T ring in flagellar motors. While MotY potentially plays an important role in motor torque generation, its impact on motor output dynamics remains poorly understood. In this study, we investigate the role of MotY in P. aeruginosa, elucidating its interactions with the two sets of stator units (MotAB and MotCD) using Förster resonance energy transfer (FRET) assays. Employing a newly developed bead assay, we characterize the dynamic behavior of flagellar motors in motY mutants, identifying MotY as the key functional protein to affect the clockwise bias of naturally unbiased motors in P. aeruginosa. Our findings reveal that MotY enhances stator assembly efficiency without affecting the overall assembly of the flagellar structure. Additionally, we demonstrate that MotY is essential for maintaining motor torque and regulating switching rates. Our study highlights the physiological significance of MotY in fine-tuning flagellar motor function in complex environments.