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Infection and Immunity, April 2006, p. 2102-2114, Vol. 74, No. 4
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.4.2102-2114.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

An IncI1 Plasmid Contributes to the Adherence of the Atypical Enteroaggregative Escherichia coli Strain C1096 to Cultured Cells and Abiotic Surfaces

Edward G. Dudley,1 Cecilia Abe,5,{dagger} Jean-Marc Ghigo,6 Patricia Latour-Lambert,6 Juan Carlos Hormazabal,7 and James P. Nataro1,2,3,4*

Center for Vaccine Development,1 Departments of Pediatrics,2 Medicine,3 Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, Maryland,4 Universidade Federal de Sao Paulo, Sao Paulo, Brazil,5 Groupe de Génétique des Biofilms, Institut Pasteur, URA CNRS 2172, 25 rue du Docteur Roux, 75724 Paris Cedex 15, France,6 Bacteriology Section, Instituto de Salud Publica, Santiago, Chile7

Received 13 September 2005/ Returned for modification 25 October 2005/ Accepted 21 December 2005

Enteroaggregative Escherichia coli (EAEC) is defined by a characteristic "stacked-brick" aggregative adherence (AA) pattern to cultured cells. In well-studied EAEC prototype strains (called typical EAEC strains), the AA phenotype requires aggregative adherence fimbriae (AAFs). However, previous studies suggest that known AAF alleles are not found in all EAEC strains. To define mechanisms contributing to adherence in an atypical strain, we studied EAEC strain C1096. An E. coli K12 derivative carrying two plasmids, designated pSERB1 and pSERB2, from C1096 adhered to cell lines and exhibited an AA pattern. Nucleotide sequence analysis of pSERB1 indicated that it is related to plasmids of the IncI1 incompatibility group. These plasmids encode genes involved in pilus-mediated conjugal transfer, as well as pilL-V, which encodes a second pilus of the type IV family. Insertional inactivation of the gene predicted to encode the major type IV pilin subunit (pilS) reduced conjugal transfer of the plasmid by 4 orders of magnitude. Adherence of the mutant strain to polystyrene and to HT29 cells was reduced by approximately 21% and 75%, respectively. In a continuous-flow microfermentor, the pilS inactivation reduced mature biofilm formation on a glass slide by approximately 50%. In addition, the simultaneous presence of both pSERB1 and pSERB2 plasmids promoted pilS-independent biofilm formation. We conclude that the IncI1 plasmid of EAEC C1096 encodes a type IV pilus that contributes to plasmid conjugation, epithelial cell adherence, and adherence to abiotic surfaces. We also observe that AA can be mediated by factors distinct from AAF adhesins.

* Corresponding author. Mailing address: University of Maryland, 685 W. Baltimore St., Baltimore, MD 21201. Phone: (410) 706 5328. Fax: (410) 706 6205. E-mail: jnataro{at}medicine.umaryland.edu.

Editor: A. D. O'Brien

{dagger} Present address: Institute of Child Health, University of Birmingham, B4 6LH Birmingham, United Kingdom.

Infection and Immunity, April 2006, p. 2102-2114, Vol. 74, No. 4
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.4.2102-2114.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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