The role in flagellar rod assembly of the N-terminal domain of Salmonella FlgJ, a flagellum-specific muramidase

J Mol Biol. 2001 Sep 14;312(2):359-69. doi: 10.1006/jmbi.2001.4963.

Abstract

The C-terminal half of the Salmonella flagellar protein FlgJ has peptidoglycan hydrolyzing activity and it has been suggested that it is a flagellum-specific muramidase which locally digests the peptidoglycan layer to permit assembly of the rod structure to proceed through the periplasmic space. It was also suggested that FlgJ might be involved in rod formation itself, although there was no direct evidence for this. We purified basal body structures from SJW1437(flgJ) transformed with plasmids encoding various mutant FlgJ proteins and found that these basal bodies possessed the periplasmic P ring but lacked the outer membrane L ring; they also lacked a hook at their distal end. All of these mutant FlgJ proteins had an altered or missing C-terminal domain but had at least the first 151 amino acid residues of the N-terminal domain. Immunoblotting analysis of fractionated cell extracts revealed that a rod/hook export class protein, FlgD, was exported to the periplasm but not to the culture supernatant in these mutants. FlgJ was shown to physically interact with several proteins, and especially FliE and FlgB, which are believed to reside at the cell-proximal end of the rod. On the basis of these results, we conclude that the N-terminal 151 amino acid residues of FlgJ are directly involved in rod formation and that the muramidase activity of FlgJ, though needed for formation of the L ring and subsequent events such as hook formation, is not essential for rod or P ring formation. In contrast, muramidase activity alone does not support rod assembly.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacterial Proteins / metabolism
  • Blotting, Western
  • Chromatography, Affinity
  • Flagella / enzymology
  • Flagella / metabolism*
  • Genetic Complementation Test
  • Multienzyme Complexes / chemistry
  • Multienzyme Complexes / deficiency
  • Multienzyme Complexes / genetics
  • Multienzyme Complexes / metabolism
  • Muramidase / chemistry*
  • Muramidase / deficiency
  • Muramidase / genetics
  • Muramidase / metabolism*
  • Mutation
  • Protein Binding
  • Protein Structure, Tertiary
  • Protein Transport
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Salmonella / cytology
  • Salmonella / enzymology*
  • Salmonella / genetics

Substances

  • Bacterial Proteins
  • FlgD protein, Salmonella typhimurium
  • Multienzyme Complexes
  • Recombinant Fusion Proteins
  • Muramidase