The Salmonella FlgA protein, a putativeve periplasmic chaperone essential for flagellar P ring formation.
P ring is a periplasmic substructure of the flagellar basal body and is believed to connect with the peptidoglycan layer in Salmonella. Two flagellar genes, flgA and flgI, are known to be indispensable for P ring formation. The flgI gene encodes the component protein of the P ring. However, the role of the flgA gene product in P ring assembly remained unknown. Here, evidence is presented that FlgA is synthesized as a precursor form and exported via the Sec secretory pathway into the periplasmic space where P ring formation takes place. Overproduction of the FlgI protein led flgA mutants to form flagella with a P ring, suggesting that FlgA plays an auxiliary role in P ring assembly. Far-Western blot analysis revealed that FlgA binds in vitro to both FlgI and FlgA itself. Though a direct FlgI-FlgI interaction in the absence of FlgA could not be demonstrated, an indirect or direct interaction between the FlgI proteins was observed in the presence of FlgA. FlgA alone was very unstable in vivo, but co-expression with FlgI could stabilize FlgA. This suggests the presence of FlgA-FlgI interaction in vivo. On the basis of these results, a hypothesis is proposed that FlgA acts as a periplasmic chaperone, which assists a polymerization reaction of FlgI into the P ring through FlgA-FlgI interaction.
Nambu
Takayuki
T
Faculty of Applied Biological Science, Hiroshima University, Kagamiyama 1-4-4, Higashi-Hiroshima, Hiroshima 739-8528, Japan1.
Kutsukake
Kazuhiro
K
Faculty of Applied Biological Science, Hiroshima University, Kagamiyama 1-4-4, Higashi-Hiroshima, Hiroshima 739-8528, Japan1.
D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D005407:Flagella / Q000737:chemistry / Q000235:genetics*; D015151:Immunoblotting; D018832:Molecular Chaperones / Q000235:genetics*; D009154:Mutation / Q000187:drug effects; D009841:Oligonucleotides; D010957:Plasmids; D011498:Protein Precursors / Q000235:genetics; D012475:Salmonella / Q000737:chemistry / Q000235:genetics*
D001426:Bacterial Proteins; C087497:FlgA protein, Salmonella; D018832:Molecular Chaperones; D009841:Oligonucleotides; D011498:Protein Precursors; C074899:flgI protein, Bacteria