Handover mechanism of the growing pilus by the bacterial outer-membrane usher FimD

Nature. 2018 Oct;562(7727):444-447. doi: 10.1038/s41586-018-0587-z. Epub 2018 Oct 3.

Abstract

Pathogenic bacteria such as Escherichia coli assemble surface structures termed pili, or fimbriae, to mediate binding to host-cell receptors1. Type 1 pili are assembled via the conserved chaperone-usher pathway2-5. The outer-membrane usher FimD recruits pilus subunits bound by the chaperone FimC via the periplasmic N-terminal domain of the usher. Subunit translocation through the β-barrel channel of the usher occurs at the two C-terminal domains (which we label CTD1 and CTD2) of this protein. How the chaperone-subunit complex bound to the N-terminal domain is handed over to the C-terminal domains, as well as the timing of subunit polymerization into the growing pilus, have previously been unclear. Here we use cryo-electron microscopy to capture a pilus assembly intermediate (FimD-FimC-FimF-FimG-FimH) in a conformation in which FimD is in the process of handing over the chaperone-bound end of the growing pilus to the C-terminal domains. In this structure, FimF has already polymerized with FimG, and the N-terminal domain of FimD swings over to bind CTD2; the N-terminal domain maintains contact with FimC-FimF, while at the same time permitting access to the C-terminal domains. FimD has an intrinsically disordered N-terminal tail that precedes the N-terminal domain. This N-terminal tail folds into a helical motif upon recruiting the FimC-subunit complex, but reorganizes into a loop to bind CTD2 during handover. Because both the N-terminal and C-terminal domains of FimD are bound to the end of the growing pilus, the structure further suggests a mechanism for stabilizing the assembly intermediate to prevent the pilus fibre diffusing away during the incorporation of thousands of subunits.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adhesins, Escherichia coli / chemistry
  • Adhesins, Escherichia coli / metabolism
  • Adhesins, Escherichia coli / ultrastructure
  • Cryoelectron Microscopy*
  • Escherichia coli / chemistry
  • Escherichia coli / metabolism*
  • Escherichia coli / ultrastructure*
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / metabolism*
  • Escherichia coli Proteins / ultrastructure*
  • Fimbriae Proteins / chemistry
  • Fimbriae Proteins / metabolism*
  • Fimbriae Proteins / ultrastructure*
  • Fimbriae, Bacterial / chemistry
  • Fimbriae, Bacterial / metabolism*
  • Fimbriae, Bacterial / ultrastructure*
  • Models, Molecular
  • Molecular Chaperones / metabolism
  • Protein Binding
  • Protein Domains
  • Protein Stability
  • Protein Subunits / chemistry
  • Protein Subunits / metabolism

Substances

  • Adhesins, Escherichia coli
  • Escherichia coli Proteins
  • FimF protein, E coli
  • FimG protein, E coli
  • Molecular Chaperones
  • Protein Subunits
  • fimC protein, E coli
  • fimD protein, E coli
  • fimH protein, E coli
  • Fimbriae Proteins