Localization of the Salmonella typhimurium Flagellar Switch Protein FliG to the Cytoplasmic M-Ring Face of the Basal Body

• Published in: Proceedings of the National Academy of Sciences - PNAS 1992-07, Vol.89 (14), p.6304-6308
• Main Authors: Francis, Noreen R., Irikura, Vera M., Yamaguchi, Shigeru, DeRosier, David J., Macnab, Robert M.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amino acids; Antibodies; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Base Sequence; Biological and medical sciences; Cell membranes; Cells; Chromosome Deletion; Electron microscopy; Flagella; Flagella - ultrastructure; Fundamental and applied biological sciences. Psychology; Genetic mutation; Genetics; Membrane Proteins; Microbiology; Microscopy, Electron; Molecular Sequence Data; Motility, taxis; Movement; Mutation; Operator regions; Proteins; Rotation; Salmonella typhimurium; Salmonella typhimurium - ultrastructure
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.89.14.6304

The direction of rotation of the bacterial flagellum is determined by the flagellar switch. We have localized FliG, one of the switch proteins of Salmonella typhimurium, to the cytoplasmic face of the M ring of the flagellar basal body. This localization was made possible by the discovery of two spontaneous mutants in which the fliF (M ring) and fliG (switch) genes were fused in-frame. In the first mutant, a deletion of 7 base pairs at the 3' end of fliF resulted in an essentially full-length fusion protein. In the second mutant, a larger deletion resulted in a fusion in which 56 amino acids from the carboxyl terminus of FliF and 94 amino acids from the amino terminus of FliG were lost. Both strains were motile and underwent switching; the first strain had a clockwise bias, and the second strain had a counterclockwise bias. Gel electrophoresis and immunoblotting of isolated hook-basal-body complexes verified that they contained the fusion proteins. Electron microscopy revealed additional mass at the cytoplasmic face of the M ring, which could be decorated with anti-FliG antibody. We conclude that the natural location for FliG is at the cytoplasmic face of the M ring and that the stoichiometric ratio between FliF and FliG in wild-type cells is probably 1:1.