Functional analysis of the flagellar genes in the fliD operon of Salmonella typhimurium.
The fliD genes of Salmonella typhimurium and Escherichia coli encode the filament-cap protein of the flagellar apparatus, which facilitates the polymerization of endogenous flagellin at the tips of the growing filaments. Previous sequence analysis of this operon in both organisms has revealed that the fliD gene constitutes an operon together with two additional genes, fliS and fliT. Based on the gene-disruption experiment in E. coli, both the fliS and fliT genes have been postulated to be necessary for flagellation. In the present study, we constructed S. typhimurium mutants in which either fliS or fliT on the chromosome was specifically disrupted. Both mutants were found to produce functional flagella, indicating that these genes are dispensable for motility development in S. typhimurium. However, flagellar filaments produced by the fliS mutant were much shorter than those produced by the wild-type strain. This indicates that the fliS mutation affects the elongation step of filament assembly. The excretion efficiency of flagellin was examined in the fliD-mutant background, where the exported flagellin molecules cannot assemble onto the hooks, resulting in their excretion into the culture media. We found that the amount of flagellin excreted was much reduced by the fliS mutation. Based on these results, we conclude that FliS facilitates the export of flagellin through the flagellum-specific export pathway.
Yokoseki
T
T
Faculty of Applied Biological Science, Hiroshima University, Japan.
Kutsukake
K
K
Ohnishi
K
K
Iino
T
T
D001426:Bacterial Proteins / Q000235:genetics* / Q000502:physiology; D001692:Biological Transport; D015153:Blotting, Western; D002465:Cell Movement; D005407:Flagella / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D005408:Flagellin / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D005798:Genes, Bacterial; D008854:Microscopy, Electron; D018832:Molecular Chaperones; D016254:Mutagenesis, Insertional; D009876:Operon; D015183:Restriction Mapping; D012486:Salmonella typhimurium / Q000235:genetics* / Q000378:metabolism
D001426:Bacterial Proteins; D018832:Molecular Chaperones; D005408:Flagellin; C076733:FlaD protein, Bacteria; C076734:FliS protein, Bacteria; C076735:fliT protein, Bacteria