Structural basis of host recognition and biofilm formation by Saf pili.

Pili are critical in host recognition， colonization and biofilm formation during bacterial infection. Here， we report the crystal structures of SafD- and SafD-SafA-SafA (SafDAA-) in Saf pili. Cell adherence assays show that SafD and SafA are both required for host recognition， suggesting a poly-adhesive mechanism for Saf pili. Moreover， the SafDAA- structure， as well as SAXS characterization， reveals an unexpected inter-molecular oligomerization， prompting the investigation of Saf-driven self-association in biofilm formation. The bead/cell aggregation and biofilm formation assays are used to demonstrate the novel function of Saf pili. Structure-based mutants targeting the inter-molecular hydrogen bonds and complementary architecture/surfaces in SafDAA- dimers significantly impaired the Saf self-association activity and biofilm formation. In summary， our results identify two novel functions of Saf pili: the poly-adhesive and self-associating activities. More importantly， Saf-Saf structures and functional characterizations help to define a pili-mediated inter-cellular oligomerizaiton mechanism for bacterial aggregation， colonization and ultimate biofilm formation.