Role of FliJ in flagellar protein export in Salmonella.
We isolated and characterized spontaneous mutants with defects in the 147-amino-acid Salmonella protein FliJ, which is a cytoplasmic component of the type III flagellar export apparatus. These mutants, including ones with null mutations, have the ability to form swarms on motility agar plates after prolonged incubation at 30 degrees C; i.e., they display a leaky motile phenotype. One mutant, SJW277, which formed significantly bigger swarms than the others, encoded only the N-terminal 73 amino acids of FliJ, one-half of the protein. At 30 degrees C, overproduction of this mutant protein improved, to wild-type levels, both motility and the ability to export both rod/hook-type (FlgD; hook capping protein) and filament-type (FliC; flagellin) substrates. At 42 degrees C, however, export was inhibited, indicating that the mutant FliJ protein was temperature sensitive. Taking advantage of this, we performed temperature upshift experiments, which demonstrated that FliJ is directly required for the export of FliC. Co-overproduction of FliJ and either of two export substrates, FliE or FlgG, hindered their aggregation in the cytoplasm. We conclude that FliJ is a general component of the flagellar export apparatus and has a chaperone-like activity for both rod/hook-type and filament-type substrates.
Minamino
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Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520-8114, USA.
Chu
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Yamaguchi
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Macnab
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RM
D019943:Amino Acid Substitution; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D001693:Biological Transport, Active; D003593:Cytoplasm / Q000378:metabolism; D005407:Flagella / Q000378:metabolism*; D005408:Flagellin / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D010641:Phenotype; D012475:Salmonella / Q000378:metabolism*; D013696:Temperature
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; C088720:FlgD protein, Salmonella typhimurium; C080688:FlhA protein, Bacteria; D008565:Membrane Proteins; C117211:flgFG protein, bacteria; D005408:Flagellin; C069253:fliJ protein, Bacteria