This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
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 Fleckenstein, J. M.
Right arrow Articles by Burkitt, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fleckenstein, J. M.
Right arrow Articles by Burkitt, M.

 Previous Article  |  Next Article 

Infection and Immunity, April 2006, p. 2245-2258, Vol. 74, No. 4
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.4.2245-2258.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Identification of a Two-Partner Secretion Locus of Enterotoxigenic Escherichia coli {dagger}

James M. Fleckenstein,1,4,5* Koushik Roy,4 Julia F. Fischer,2,3 and Michael Burkitt3

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

Received 7 November 2005/ Returned for modification 21 December 2005/ Accepted 10 January 2006

Enterotoxigenic Escherichia coli (ETEC) remains a formidable cause of diarrheal illness worldwide. At present, there is no vaccine that provides broad-based protection against ETEC. A 'phoA-based self-cloning mutagenesis system, TnphoA.ts, employed to identify novel ETEC surface antigens, led to identification of an ETEC two-partner secretion locus (etpBAC) on the pCS1 virulence plasmid of prototype strain H10407. Cloning and expression of etpBAC in recombinant E. coli LMG194(pJY019) resulted in secretion of a high-molecular-weight (HMW) glycosylated exoprotein. This glycoprotein, EtpA, exhibits linear peptide sequence and predicted structural homologies with known HMW adhesins produced by other two-partner secretion loci. Antibodies directed against recombinant EtpA (anti-rEtpA.6H) recognized an HMW protein in culture supernatants of ETEC strains H10407 and LMG194(pJY019) but not in culture supernatant of strain H10407-P, which lacks the 92-kb pCS1 plasmid, or an isogenic etpA mutant. etpA mutants were deficient in adherence to intestinal epithelial cells in vitro, and anti-rEtpA.6H antibodies inhibited association of H10407 with target epithelial cells. Cloning and expression of etpB in recombinant E. coli were sufficient to confer adherence. Screening of multiple ETEC isolates for the etpBAC locus by colony hybridization and by EtpA immunoblotting suggested that EtpA is one of the most common antigens secreted by these pathogens. Together, these results indicate that the newly identified ETEC two-partner secretion locus directs the secretion of a high-molecular-weight glycosylated protein, EtpA, that in concert with the putative EtpB transporter participates in adherence of H10407 to epithelial cells, thereby expanding the repertoire of potential ETEC virulence proteins and vaccine candidates.

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

Editor: A. D. O'Brien

{dagger} Supplemental material for this article may be found at http://iai.asm.org/.

Infection and Immunity, April 2006, p. 2245-2258, Vol. 74, No. 4
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.4.2245-2258.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Neil, R. B., Apicella, M. A. (2009). Role of HrpA in Biofilm Formation of Neisseria meningitidis and Regulation of the hrpBAS Transcripts. Infect. Immun. 77: 2285-2293 [Abstract] [Full Text]  
  • Shaheen, H. I., Abdel Messih, I. A., Klena, J. D., Mansour, A., El-Wakkeel, Z., Wierzba, T. F., Sanders, J. W., Khalil, S. B., Rockabrand, D. M., Monteville, M. R., Rozmajzl, P. J., Svennerholm, A. M., Frenck, R. W. (2009). Phenotypic and Genotypic Analysis of Enterotoxigenic Escherichia coli in Samples Obtained from Egyptian Children Presenting to Referral Hospitals. J. Clin. Microbiol. 47: 189-197 [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]  
  • Sjoling, A., Wiklund, G., Savarino, S. J., Cohen, D. I., Svennerholm, A.-M. (2007). Comparative Analyses of Phenotypic and Genotypic Methods for Detection of Enterotoxigenic Escherichia coli Toxins and Colonization Factors. J. Clin. Microbiol. 45: 3295-3301 [Abstract] [Full Text]  
  • Balder, R., Hassel, J., Lipski, S., Lafontaine, E. R. (2007). Moraxella catarrhalis Strain O35E Expresses Two Filamentous Hemagglutinin-Like Proteins That Mediate Adherence to Human Epithelial Cells. Infect. Immun. 75: 2765-2775 [Abstract] [Full Text]  
  • Choi, P. S., Dawson, A. J., Bernstein, H. D. (2007). Characterization of a Novel Two-Partner Secretion System in Escherichia coli O157:H7. J. Bacteriol. 189: 3452-3461 [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»