Analysis of the requirements for pilus biogenesis at the outer membrane usher and the function of the usher C‐terminus

Summary Uropathogenic strains of Escherichia coli assemble type 1 and P pili to colonize the bladder and kidney respectively. These pili are prototype structures assembled by the chaperone/usher secretion pathway. In this pathway, a periplasmic chaperone works together with an outer membrane (OM) usher to control the folding of pilus subunits, their assembly into a pilus fibre and secretion of the fibre to the cell surface. The usher serves as the assembly and secretion platform in the OM. The usher has distinct functional domains, with the N‐terminus providing the initial targeting site for chaperone–subunit complexes and the C‐terminus required for subsequent stages of pilus biogenesis. In this study, we investigated the molecular interactions occurring at the usher during pilus biogenesis and the function of the usher C‐terminus. We provide genetic and biochemical evidence that the usher functions as a complex in the OM and that interaction of the pilus adhesin with the usher is critical to prime the usher for pilus biogenesis. Analysis of C‐terminal truncation and substitution mutants of the P pilus usher PapC demonstrated that the C‐terminus is required for proper binding of chaperone–subunit complexes to the usher and plays an important role in assembly of complete pili.