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 Patel, S. K.
Right arrow Articles by Fleckenstein, J. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Patel, S. K.
Right arrow Articles by Fleckenstein, J. M.

 Previous Article  |  Next Article 

Infection and Immunity, March 2004, p. 1786-1794, Vol. 72, No. 3
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.3.1786-1794.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Identification and Molecular Characterization of EatA, an Autotransporter Protein of Enterotoxigenic Escherichia coli

Seema K. Patel,1 Jimmie Dotson,2 Kenneth P. Allen,3 and James M. Fleckenstein1,4,5*

Departments of Medicine,1 Comparative Medicine,3 Molecular Sciences,4 College of Medicine, University of Tennessee Health Science Center,2 Medicine Service, Veterans Affairs Medical Center, Memphis, Tennessee5

Received 15 July 2003/ Returned for modification 16 September 2003/ Accepted 12 November 2003

Enterotoxigenic Escherichia coli (ETEC) strains remain a formidable cause of diarrheal disease. To identify novel surface proteins of ETEC, we performed TnphoA mutagenesis of prototype ETEC strain H10407 and discovered a secreted protein not previously recognized in ETEC. DNA sequencing of the interrupted locus in mutant TnphoA.977 revealed a candidate 4,095-bp open reading frame without significant homology to commensal E. coli K-12 genomic DNA. Translation of this sequence revealed that it encoded a predicted peptide of 147.7 kDa that bears significant homology to members of the autotransporter family of bacterial virulence factors, particularly the serine protease autotransporters of the Enterobacteriaceae proteins. The gene identified in H10407, eatA (ETEC autotransporter A), encodes a potential serine protease motif (GDSGSP) in the secreted amino-terminal domain, and the predicted peptide shows more than 80% homology with SepA, a virulence protein secreted by Shigella flexneri. DNA hybridization and PCR demonstrated that eatA resides on the 92-kDa pCS1 virulence plasmid of H10407 and that it is present in multiple clinical ETEC strains. Immunoblots with antisera directed against a recombinant EatA passenger protein fragment identified a 110-kDa protein in supernatants purified from H10407 but not from the TnphoA.977 mutant or H10407-P, which lacks pCS1. EatA possesses serine protease activity that is abolished by mutations within a serine protease catalytic triad formed by residues H134, D162, and S267. Finally, interruption of the eatA gene retarded fluid accumulation in the rabbit ileal loop model, suggesting that this autotransporter contributes to the virulence of ETEC.

* Corresponding author. Mailing address: Research Service (151), Veterans Affairs Medical Center, 1030 Jefferson Ave., Memphis, TN 38104. Phone: (901) 448-5786. Fax: (901) 577-7273. E-mail: jflecke1{at}tennessee.edu.

Editor: B. B. Finlay

Infection and Immunity, March 2004, p. 1786-1794, Vol. 72, No. 3
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.3.1786-1794.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Betancourt-Sanchez, M., Navarro-Garcia, F. (2009). Pet secretion, internalization and induction of cell death during infection of epithelial cells by enteroaggregative Escherichia coli. Microbiology 155: 2895-2906 [Abstract] [Full Text]  
  • Hu, Y.-h., Liu, C.-s., Hou, J.-h., Sun, L. (2009). Identification, Characterization, and Molecular Application of a Virulence-Associated Autotransporter from a Pathogenic Pseudomonas fluorescens Strain. Appl. Environ. Microbiol. 75: 4333-4340 [Abstract] [Full Text]  
  • Rasko, D. A., Rosovitz, M. J., Myers, G. S. A., Mongodin, E. F., Fricke, W. F., Gajer, P., Crabtree, J., Sebaihia, M., Thomson, N. R., Chaudhuri, R., Henderson, I. R., Sperandio, V., Ravel, J. (2008). The Pangenome Structure of Escherichia coli: Comparative Genomic Analysis of E. coli Commensal and Pathogenic Isolates. J. Bacteriol. 190: 6881-6893 [Abstract] [Full Text]  
  • Roy, K., Hamilton, D., Allen, K. P., Randolph, M. P., Fleckenstein, J. M. (2008). The EtpA Exoprotein of Enterotoxigenic Escherichia coli Promotes Intestinal Colonization and Is a Protective Antigen in an Experimental Model of Murine Infection. Infect. Immun. 76: 2106-2112 [Abstract] [Full Text]  
  • Restieri, C., Garriss, G., Locas, M.-C., Dozois, C. M. (2007). Autotransporter-Encoding Sequences Are Phylogenetically Distributed among Escherichia coli Clinical Isolates and Reference Strains. Appl. Environ. Microbiol. 73: 1553-1562 [Abstract] [Full Text]  
  • Coster, T. S., Wolf, M. K., Hall, E. R., Cassels, F. J., Taylor, D. N., Liu, C. T., Trespalacios, F. C., DeLorimier, A., Angleberger, D. R., McQueen, C. E. (2007). Immune Response, Ciprofloxacin Activity, and Gender Differences after Human Experimental Challenge by Two Strains of Enterotoxigenic Escherichia coli. Infect. Immun. 75: 252-259 [Abstract] [Full Text]  
  • Turner, S. M., Chaudhuri, R. R., Jiang, Z.-D., DuPont, H., Gyles, C., Penn, C. W., Pallen, M. J., Henderson, I. R. (2006). Phylogenetic Comparisons Reveal Multiple Acquisitions of the Toxin Genes by Enterotoxigenic Escherichia coli Strains of Different Evolutionary Lineages. J. Clin. Microbiol. 44: 4528-4536 [Abstract] [Full Text]  
  • Maroncle, N. M., Sivick, K. E., Brady, R., Stokes, F.-E., Mobley, H. L. T. (2006). Protease Activity, Secretion, Cell Entry, Cytotoxicity, and Cellular Targets of Secreted Autotransporter Toxin of Uropathogenic Escherichia coli. Infect. Immun. 74: 6124-6134 [Abstract] [Full Text]  
  • Fleckenstein, J. M., Roy, K., Fischer, J. F., Burkitt, M. (2006). Identification of a Two-Partner Secretion Locus of Enterotoxigenic Escherichia coli. Infect. Immun. 74: 2245-2258 [Abstract] [Full Text]  
  • Chen, Q., Savarino, S. J., Venkatesan, M. M. (2006). Subtractive hybridization and optical mapping of the enterotoxigenic Escherichia coli H10407 chromosome: isolation of unique sequences and demonstration of significant similarity to the chromosome of E. coli K-12.. Microbiology 152: 1041-1054 [Abstract] [Full Text]  
  • Allen, K. P., Randolph, M. M., Fleckenstein, J. M. (2006). Importance of Heat-Labile Enterotoxin in Colonization of the Adult Mouse Small Intestine by Human Enterotoxigenic Escherichia coli Strains. Infect. Immun. 74: 869-875 [Abstract] [Full Text]  
  • Froehlich, B., Parkhill, J., Sanders, M., Quail, M. A., Scott, J. R. (2005). The pCoo Plasmid of Enterotoxigenic Escherichia coli Is a Mosaic Cointegrate. J. Bacteriol. 187: 6509-6516 [Abstract] [Full Text]  
  • Parham, N. J., Pollard, S. J., Desvaux, M., Scott-Tucker, A., Liu, C., Fivian, A., Henderson, I. R. (2005). Distribution of the Serine Protease Autotransporters of the Enterobacteriaceae among Extraintestinal Clinical Isolates of Escherichia coli. J. Clin. Microbiol. 43: 4076-4082 [Abstract] [Full Text]  
  • Henderson, I. R., Navarro-Garcia, F., Desvaux, M., Fernandez, R. C., Ala'Aldeen, D. (2004). Type V Protein Secretion Pathway: the Autotransporter Story. Microbiol. Mol. Biol. Rev. 68: 692-744 [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»