Rapid cytoskeletal response of epithelial cells to force generation by type IV pili

Many bacterial pathogens interfere with cellular functions including phagocytosis and barrier integrity. The human pathogen Neissieria gonorrhoeae generates grappling hooks for adhesion, spreading, and induction of signal cascades that lead to formation cortical plaques containing f-actin and ezrin. It is unclear whether high mechanical forces generated by type IV pili (T4P) are a direct signal that leads to cytoskeletal rearrangements and at which time scale the cytoskeletal response occurs. Here we used laser tweezers to mimic type IV pilus mediated force generation by T4P-coated beads on the order of 100 pN. We found that actin-EGFP and ezrin-EGFP accumulated below pilus-coated beads when force was applied. Within 2 min, accumulation significantly exceeded controls without force or without pili, demonstrating that T4P-generated force rapidly induces accumulation of plaque proteins. This finding adds mechanical force to the many strategies by which bacteria modulate the host cell cytoskeleton.