FrzS acts as a polar beacon to recruit SgmX, a central activator of type IV pili during Myxococcus xanthus motility

In rod‐shaped bacteria, type IV pili (Tfp) promote twitching motility by assembling and retracting at the cell pole. In Myxococcus xanthus , a bacterium that moves in highly coordinated cell groups, Tfp are activated by a polar activator protein, SgmX. However, while it is known that the Ras‐like protein MglA is required for unipolar targeting, how SgmX accesses the cell pole to activate Tfp is unknown. Here, we demonstrate that a polar beacon protein, FrzS, recruits SgmX at the cell pole. We identified two main functional domains, including a Tfp‐activating domain and a polar‐binding domain. Within the latter, we show that the direct binding of MglA‐GTP unveils a hidden motif that binds directly to the FrzS N‐terminal response regulator (CheY). Structural analyses reveal that this binding occurs through a novel binding interface for response regulator domains. In conclusion, the findings unveil the protein interaction network leading to the spatial activation of Tfp at the cell pole. This tripartite system is at the root of complex collective behaviours in this predatory bacterium. Synopsis In Myxococcus xanthus , social motility depends on activation of Type IV Pili by the polar protein SgmX. Here, the cell pole‐localized protein FrzS is shown to mediate SgmX polar recruitment via an atypical protein‐protein interaction. FrzS recruits the Type IV Pili‐activating protein SgmX to the cell pole. SgmX interacts with FrzSCheY domain when complexed with MglA‐GTP. The atypical response regulator (CheY) domain of FrzS has acquired a protein‐protein interaction function. Structural analyses show that FrzSCheY and SgmX interact via a novel binding interface of response regulator domains. Graphical Abstract Mutational analysis reveals a new SgmX motif mediating its recruitment to cell poles via the social motility regulator protein FrzS.