Abstract
Pseudomonas aeruginosa type IV pili (TFP) are important for twitching motility and biofilm formation. TFP have been implicated in surface sensing, a process whereby surface-engaged cells upregulate the synthesis of the second messenger cAMP to propagate a signaling cascade leading to biofilm initiation and repression of motility. Here, we showed that mutations in PilA impairing proteolytic processing of the prepilin into mature pilin as well as the disruption of essential TFP components, including the PilC platform protein and PilB assembly motor protein, fail to induce surface-dependent cAMP signaling. We showed that TFP retraction by surface-engaged cells was required to induce signaling and that the retractile motor PilT was both necessary and sufficient to power surface-specific induction of cAMP. Furthermore, full TFP function required to support twitching motility is not required for robust cAMP signalling. The PilU retraction motor, in contrast, was unable to support full signaling in the absence of PilT. Finally, while we confirmed that PilA and PilJ interacted by bacterial two-hybrid analysis, our data do not support the current model that PilJ-PilA interaction drives cAMP signaling. IMPORTANCE Surface sensing by P. aeruginosa requires TFP. TFP plays a critical role in the induction of the second messenger cAMP upon surface contact; this second messenger is part of a larger cascade involved in the transition from a planktonic to a biofilm lifestyle. Here, we showed that TFP must be deployed and actively retracted by the PilT motor for the full induction of cAMP signaling. Furthermore, the mechanism whereby TFP retraction triggers cAMP induction is not well understood, and our data argue against one of the current models in the field proposed to address this knowledge gap.