Genetic and biochemical analysis of Salmonella typhimurium FliI, a flagellar protein related to the catalytic subunit of the F0F1 ATPase and to virulence proteins of mammalian and plant pathogens.
FliI is a Salmonella typhimurium protein that is needed for flagellar assembly and may be involved in a specialized protein export pathway that proceeds without signal peptide cleavage. FliI shows extensive sequence similarity to the catalytic beta subunit of the F0F1 ATPase (A. P. Volger, M. Homma, V. M. Irikura, and R. M. Macnab, J. Bacteriol. 173:3564-3572, 1991). It is even more similar to the Spa47 protein of Shigella flexneri (M. M. Venkatesan, J. M. Buysse, and E. V. Oaks, J. Bacteriol. 174:1990-2001, 1992) and the HrpB6 protein of Xanthomonas campestris (S. Fenselau, I. Balbo, and U. Bonas, Mol. Plant-Microbe Interact. 5:390-396, 1992), which are believed to play a role in the export of virulence proteins. Site-directed mutagenesis of residues in FliI that correspond to catalytically important residues in the F1 beta subunit resulted in loss of flagellation, supporting the hypothesis that FliI is an ATPase. FliI was overproduced and purified almost to homogeneity. It demonstrated ATP binding but not hydrolysis. An antibody raised against FliI permitted detection of the protein in wild-type cells and an estimate of about 1,500 subunits per cell. An antibody directed against the F1 beta subunit of Escherichia coli cross-reacted with FliI, confirming that the proteins are structurally related. The relationship between three proteins involved in flagellar assembly (FliI, FlhA, and FliP) and homologs in a variety of virulence systems is discussed.
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06511-8148.
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Dreyfus
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Williams
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Kawagishi
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Macnab
D000255:Adenosine Triphosphate / Q000378:metabolism*; D001419:Bacteria / Q000472:pathogenicity; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D001483:Base Sequence; D003429:Cross Reactions; D005407:Flagella / Q000502:physiology*; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D010935:Plant Diseases; D011506:Proteins; D006180:Proton-Translocating ATPases / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D012486:Salmonella typhimurium / Q000235:genetics* / Q000472:pathogenicity; D014774:Virulence; D037521:Virulence Factors
D001426:Bacterial Proteins; D011506:Proteins; D011994:Recombinant Proteins; D037521:Virulence Factors; C069251:fliI protein, bacteria; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases