FliN is a major structural protein of the C-ring in the Salmonella typhimurium flagellar basal body

Rongbao Zhao, Naveen Pathak, Howard Jaffe, Thomas S. Reese, Shahid Khan

Research output: Contribution to journalArticle

84 Citations (Scopus)

Abstract

The Salmonella typhimurium FliN protein has been proposed to form a mutually interacting complex with FliG and FliM, the switch complex, that is required for flagellar morphogenesis and function. We have used affinity chromatography for purification of extended flagellar basal bodies sufficient for quantitative analysis of their protein composition. The belied, extended structure is predominantly comprised of the switch complex proteins; with FliN present in the most copies (111 ± 13). This explains why single, missense fliN, fliG or fliM mutations, found in many non-motile strains, can alter the belied morphology. Cell lysates from these strains contained the wild-type complement of FliG, FliM and FliN; but the basal bodies lacked the outer, cytoplasmic(C)-ring of the bell and were separated by sedimentation from FliM and FliN. The amount of FliG present in basal bodies from wild-type and one such mutant, FliN100LP, was comparable. These data show that: (1) the mutations define a FliG and FliMfliN multiple contact interface important for motility. (2) FliG is responsible for the increased size of the membrane-embedded MS-ring complex of belied relative to acid-treated basal bodies. (3) FliN, together with FliM, account for most of the C-ring. As a major component of the C-ring, FliN is distinct from the other proteins implicated in axial flagellar protein export. Inner, cytoplasmic rod basal substructure, seen by negative-stain and quick-freeze replica electron microscopy, may gate such export. Lack of connectivity between the cytoplasmic rod and ring substructures places contacts between FliG and FliMFliN at the periphery of the basal body, proximal to the flagellar intramembrane ring particles. This topology is consistent with models where torque results from interaction of circumferential arrays of the switch complex proteins with the ring particles.

Original languageEnglish (US)
Pages (from-to)195-208
Number of pages14
JournalJournal of Molecular Biology
Volume261
Issue number2
DOIs
StatePublished - Aug 16 1996

Fingerprint

Basal Bodies
Salmonella typhimurium
Protein C
Switch Genes
Proteins
Somatotypes
Mutation
Torque
Morphogenesis
Affinity Chromatography
Electron Microscopy
Coloring Agents
Acids
Membranes

Keywords

  • Chromatography
  • Electron microscopy
  • Macromolecular assembly
  • Molecular motor
  • Protein export

ASJC Scopus subject areas

  • Virology

Cite this

FliN is a major structural protein of the C-ring in the Salmonella typhimurium flagellar basal body. / Zhao, Rongbao; Pathak, Naveen; Jaffe, Howard; Reese, Thomas S.; Khan, Shahid.

In: Journal of Molecular Biology, Vol. 261, No. 2, 16.08.1996, p. 195-208.

Research output: Contribution to journalArticle

Zhao, Rongbao ; Pathak, Naveen ; Jaffe, Howard ; Reese, Thomas S. ; Khan, Shahid. / FliN is a major structural protein of the C-ring in the Salmonella typhimurium flagellar basal body. In: Journal of Molecular Biology. 1996 ; Vol. 261, No. 2. pp. 195-208.
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