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
Por:
Dreyfus G., WIlliams A.W., Kawagishi I., Macnab R.M.
Publicada:
1 ene 1993
Resumen:
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 ? subunit of the F0F1 ATPase (A. P. Vogier, 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 ? 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 ? 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.
Filiaciones:
Dreyfus G.:
Molecular Biophysics/Biochem. Dept., Yale University, New Haven, CT 06511-8148, United States
WIlliams A.W.:
Molecular Biophysics/Biochem. Dept., Yale University, New Haven, CT 06511-8148, United States
Kawagishi I.:
Molecular Biophysics/Biochem. Dept., Yale University, New Haven, CT 06511-8148, United States
Macnab R.M.:
Molecular Biophysics/Biochem. Dept., Yale University, New Haven, CT 06511-8148, United States
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