title: Substrate specificity classes and the recognition signal for Salmonella type III flagellar export.
abstract: Most flagellar proteins of Salmonella are exported to their assembly destination via a specialized apparatus. This apparatus is a member of the type III superfamily, which is widely used for secretion of virulence factors by pathogenic bacteria. Extensive studies have been carried out on the export of several of the flagellar proteins, most notably the hook protein (FlgE), the hook-capping protein (FlgD), and the filament protein flagellin (FliC). This has led to the concept of two export specificity classes, the rod/hook type and the filament type. However, little direct experimental evidence has been available on the export properties of the basal-body rod proteins (FlgB, FlgC, FlgF, and FlgG), the putative MS ring-rod junction protein (FliE), or the muramidase and putative rod-capping protein (FlgJ). In this study, we have measured the amounts of these proteins exported before and after hook completion. Their amounts in the culture supernatant from a flgE mutant (which is still at the hook-type specificity stage) were much higher than those from a flgK mutant (which has advanced to the filament-type specificity stage), placing them in the same class as the hook-type proteins. Overproduction of FliE, FlgB, FlgC, FlgF, FlgG, or FlgJ caused inhibition of the motility of wild-type cells and inhibition of the export of the hook-capping protein FlgD. We also examined the question of whether export and translation are linked and found that all substrates tested could be exported after protein synthesis had been blocked by spectinomycin or chloramphenicol. We conclude that the amino acid sequence of these proteins suffices to mediate their recognition and export.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520-8114, USA.
authors.forename: Takanori
authors.initials: T
authors.lastname: Hirano
authors.forename: Tohru
authors.initials: T
authors.lastname: Minamino
authors.forename: Keiichi
authors.initials: K
authors.lastname: Namba
authors.forename: Robert M
authors.initials: RM
authors.lastname: Macnab
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D001692:Biological Transport / Q000187:drug effects; D002701:Chloramphenicol / Q000494:pharmacology; D017077:Culture Media, Conditioned; D029968:Escherichia coli Proteins; D005407:Flagella / Q000378:metabolism*; D007544:Isopropyl Thiogalactoside; D009068:Movement; D009154:Mutation; D010957:Plasmids; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012475:Salmonella / Q000187:drug effects / Q000378:metabolism*; D000198:Spectinomycin / Q000494:pharmacology; D013379:Substrate Specificity
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D017077:Culture Media, Conditioned; D029968:Escherichia coli Proteins; D011500:Protein Synthesis Inhibitors; C117211:flgFG protein, bacteria; C073764:fliE protein, E coli; D007544:Isopropyl Thiogalactoside; D002701:Chloramphenicol; D000198:Spectinomycin