This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Minamino, T.
Right arrow Articles by Macnab, R. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Minamino, T.
Right arrow Articles by Macnab, R. M.

 Previous Article  |  Next Article 

Journal of Bacteriology, September 2000, p. 4906-4914, Vol. 182, No. 17
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Domain Structure of Salmonella FlhB, a Flagellar Export Component Responsible for Substrate Specificity Switching

Tohru Minamino and Robert M. Macnab*

Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520-8114

Received 5 April 2000/Accepted 2 June 2000

We have investigated the properties of the cytoplasmic domain (FlhBC) of the 383-amino-acid Salmonella membrane protein FlhB, a component of the type III flagellar export apparatus. FlhB, along with the hook-length control protein FliK, mediates the switching of export specificity from rod- and hook-type substrates to filament-type substrates during flagellar morphogenesis. Wild-type FlhBC was unstable (half-life, ca. 5 min), being specifically cleaved at Pro-270 into two polypeptides, FlhBCN and FlhBCC, which retained the ability to interact with each other after cleavage. Full-length wild-type FlhB was also subject to cleavage. Coproduction of the cleavage products, FlhBDelta CC (i.e., the N-terminal transmembrane domain FlhBTM plus FlhBCN) and FlhBCC, resulted in restoration of both motility and flagellar protein export to an flhB mutant host, indicating that the two polypeptides were capable of productive association. Mutant FlhB proteins that can undergo switching of substrate specificity even in the absence of FliK were much more resistant to cleavage (half-lives, 20 to 60 min). The cleavage products of wild-type FlhBC, existing as a FlhBCN-FlhBCC complex on an affinity blot membrane, bound the rod- and hook-type substrate FlgD more strongly than the filament-type substrate FliC. In contrast, the intact form of FlhBC (mutant or wild type) or the FlhBCC polypeptide alone bound FlgD and FliC to about the same extent. FlhBCN by itself did not bind substrates appreciably. We propose that FlhBC has two substrate specificity states and that a conformational change, mediated by the interaction between FlhBCN and FlhBCC, is responsible for the specificity switching process. FliK itself is an export substrate; its binding properties for FlhBC resemble those of FlgD and do not provide any evidence for a physical interaction beyond that of the export process.

* Corresponding author. Mailing address: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520-8114. Phone: (203) 432-5590. Fax: (203) 432-9782. E-mail: robert.macnab{at}yale.edu.

Journal of Bacteriology, September 2000, p. 4906-4914, Vol. 182, No. 17
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Fowler, J. M., Wulff, C. R., Straley, S. C., Brubaker, R. R. (2009). Growth of calcium-blind mutants of Yersinia pestis at 37 {degrees}C in permissive Ca2+-deficient environments. Microbiology 155: 2509-2521 [Abstract] [Full Text]  
  • Bjornfot, A.-C., Lavander, M., Forsberg, A., Wolf-Watz, H. (2009). Autoproteolysis of YscU of Yersinia pseudotuberculosis Is Important for Regulation of Expression and Secretion of Yop Proteins. J. Bacteriol. 191: 4259-4267 [Abstract] [Full Text]  
  • Hirano, T., Mizuno, S., Aizawa, S.-I., Hughes, K. T. (2009). Mutations in Flk, FlgG, FlhA, and FlhE That Affect the Flagellar Type III Secretion Specificity Switch in Salmonella enterica. J. Bacteriol. 191: 3938-3949 [Abstract] [Full Text]  
  • Joslin, S. N., Hendrixson, D. R. (2009). Activation of the Campylobacter jejuni FlgSR Two-Component System Is Linked to the Flagellar Export Apparatus. J. Bacteriol. 191: 2656-2667 [Abstract] [Full Text]  
  • Smith, T. G., Pereira, L., Hoover, T. R. (2009). Helicobacter pylori FlhB processing-deficient variants affect flagellar assembly but not flagellar gene expression. Microbiology 155: 1170-1180 [Abstract] [Full Text]  
  • Yang, T.-C., Leu, Y.-W., Chang-Chien, H.-C., Hu, R.-M. (2009). Flagellar Biogenesis of Xanthomonas campestris Requires the Alternative Sigma Factors RpoN2 and FliA and Is Temporally Regulated by FlhA, FlhB, and FlgM. J. Bacteriol. 191: 2266-2275 [Abstract] [Full Text]  
  • Riordan, K. E., Sorg, J. A., Berube, B. J., Schneewind, O. (2008). Impassable YscP Substrates and Their Impact on the Yersinia enterocolitica Type III Secretion Pathway. J. Bacteriol. 190: 6204-6216 [Abstract] [Full Text]  
  • Kamal, N., Dorrell, N., Jagannathan, A., Turner, S. M., Constantinidou, C., Studholme, D. J., Marsden, G., Hinds, J., Laing, K. G., Wren, B. W., Penn, C. W. (2007). Deletion of a previously uncharacterized flagellar-hook-length control gene fliK modulates the {sigma}54-dependent regulon in Campylobacter jejuni. Microbiology 153: 3099-3111 [Abstract] [Full Text]  
  • Wand, M. E., Sockett, R. E., Evans, K. J., Doherty, N., Sharp, P. M., Hardie, K. R., Winzer, K. (2006). Helicobacter pylori FlhB Function: the FlhB C-Terminal Homologue HP1575 Acts as a "Spare Part" To Permit Flagellar Export When the HP0770 FlhBCC Domain Is Deleted.. J. Bacteriol. 188: 7531-7541 [Abstract] [Full Text]  
  • Ferris, H. U., Furukawa, Y., Minamino, T., Kroetz, M. B., Kihara, M., Namba, K., Macnab, R. M. (2005). FlhB Regulates Ordered Export of Flagellar Components via Autocleavage Mechanism. J. Biol. Chem. 280: 41236-41242 [Abstract] [Full Text]  
  • Ghosh, P. (2004). Process of Protein Transport by the Type III Secretion System. Microbiol. Mol. Biol. Rev. 68: 771-795 [Abstract] [Full Text]  
  • McMurry, J. L., Van Arnam, J. S., Kihara, M., Macnab, R. M. (2004). Analysis of the Cytoplasmic Domains of Salmonella FlhA and Interactions with Components of the Flagellar Export Machinery. J. Bacteriol. 186: 7586-7592 [Abstract] [Full Text]  
  • Van Arnam, J. S., McMurry, J. L., Kihara, M., Macnab, R. M. (2004). Analysis of an Engineered Salmonella Flagellar Fusion Protein, FliR-FlhB. J. Bacteriol. 186: 2495-2498 [Abstract] [Full Text]  
  • Journet, L., Agrain, C., Broz, P., Cornelis, G. R. (2003). The Needle Length of Bacterial Injectisomes Is Determined by a Molecular Ruler. Science 302: 1757-1760 [Abstract] [Full Text]  
  • Gauthier, A., Finlay, B. B. (2003). Translocated Intimin Receptor and Its Chaperone Interact with ATPase of the Type III Secretion Apparatus of Enteropathogenic Escherichia coli. J. Bacteriol. 185: 6747-6755 [Abstract] [Full Text]  
  • Creasey, E. A., Delahay, R. M., Daniell, S. J., Frankel, G. (2003). Yeast two-hybrid system survey of interactions between LEE-encoded proteins of enteropathogenic Escherichia coli. Microbiology 149: 2093-2106 [Abstract] [Full Text]  
  • Edqvist, P. J., Olsson, J., Lavander, M., Sundberg, L., Forsberg, A., Wolf-Watz, H., Lloyd, S. A. (2003). YscP and YscU Regulate Substrate Specificity of the Yersinia Type III Secretion System. J. Bacteriol. 185: 2259-2266 [Abstract] [Full Text]  
  • Bardy, S. L., Ng, S. Y. M., Jarrell, K. F. (2003). Prokaryotic motility structures. Microbiology 149: 295-304 [Abstract] [Full Text]  
  • Lavander, M., Sundberg, L., Edqvist, P. J., Lloyd, S. A., Wolf-Watz, H., Forsberg, A. (2002). Proteolytic Cleavage of the FlhB Homologue YscU of Yersinia pseudotuberculosis Is Essential for Bacterial Survival but Not for Type III Secretion. J. Bacteriol. 184: 4500-4509 [Abstract] [Full Text]  
  • Matz, C., van Vliet, A. H. M., Ketley, J. M., Penn, C. W. (2002). Mutational and transcriptional analysis of the Campylobacter jejuni flagellar biosynthesis gene flhB. Microbiology 148: 1679-1685 [Abstract] [Full Text]  
  • Segura, A., Duque, E., Hurtado, A., Ramos, J. L. (2001). Mutations in Genes Involved in the Flagellar Export Apparatus of the Solvent-Tolerant Pseudomonas putida DOT-T1E Strain Impair Motility and Lead to Hypersensitivity to Toluene Shocks. J. Bacteriol. 183: 4127-4133 [Abstract] [Full Text]  
  • Vallet, I., Olson, J. W., Lory, S., Lazdunski, A., Filloux, A. (2001). The chaperone/usher pathways of Pseudomonas aeruginosa: Identification of fimbrial gene clusters (cup) and their involvement in biofilm formation. Proc. Natl. Acad. Sci. USA 10.1073/pnas.111551898v1 [Abstract] [Full Text]  
  • Vallet, I., Olson, J. W., Lory, S., Lazdunski, A., Filloux, A. (2001). The chaperone/usher pathways of Pseudomonas aeruginosa: Identification of fimbrial gene clusters (cup) and their involvement in biofilm formation. Proc. Natl. Acad. Sci. USA 98: 6911-6916 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»