Co-Folding of a FliF-FliG Split Domain Forms the Basis of the MS:C Ring Interface within the Bacterial Flagellar Motor

Structure. 2017 Feb 7;25(2):317-328. doi: 10.1016/j.str.2016.12.006. Epub 2017 Jan 12.

Abstract

The interface between the membrane (MS) and cytoplasmic (C) rings of the bacterial flagellar motor couples torque generation to rotation within the membrane. The structure of the C-terminal helices of the integral membrane protein FliF (FliFC) bound to the N terminal domain of the switch complex protein FliG (FliGN) reveals that FliGN folds around FliFC to produce a topology that closely resembles both the middle and C-terminal domains of FliG. The interface is consistent with solution-state nuclear magnetic resonance, small-angle X-ray scattering, in vivo interaction studies, and cellular motility assays. Co-folding with FliFC induces substantial conformational changes in FliGN and suggests that FliF and FliG have the same stoichiometry within the rotor. Modeling the FliFC:FliGN complex into cryo-electron microscopy rotor density updates the architecture of the middle and upper switch complex and shows how domain shuffling of a conserved interaction module anchors the cytoplasmic rotor to the membrane.

Keywords: X-ray structure; assembly; chemotaxis; cross-linking; domain shuffling; flagellar motor; molecular machine; motility; switch complex.

MeSH terms

  • Amino Acid Motifs
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Binding Sites
  • Biomechanical Phenomena
  • Cell Membrane / chemistry*
  • Cell Membrane / ultrastructure
  • Cloning, Molecular
  • Crystallography, X-Ray
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Flagella / chemistry*
  • Flagella / ultrastructure
  • Gene Expression
  • Membrane Proteins / chemistry*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Models, Molecular
  • Protein Binding
  • Protein Conformation, alpha-Helical
  • Protein Folding
  • Protein Interaction Domains and Motifs
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Thermotoga maritima / chemistry*
  • Thermotoga maritima / ultrastructure

Substances

  • Bacterial Proteins
  • Flif protein, Bacteria
  • Flig protein, Bacteria
  • Membrane Proteins
  • Recombinant Fusion Proteins