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Infection and Immunity, October 2002, p. 5416-5427, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5416-5427.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Identification and Characterization of lpfABCC'DE, a Fimbrial Operon of Enterohemorrhagic Escherichia coli O157:H7

Alfredo G. Torres,1 Jorge A. Giron,1,2 Nicole T. Perna,3 Valerie Burland,4 Fred R. Blattner,4 Fabiola Avelino-Flores,2 and James B. Kaper1*

Center for Vaccine Development and Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, Maryland 21201,1 Centro de Investigaciones Microbiológicas, Instituto de Ciencias, Benemérita Universidad Autónoma de Puebla, Puebla, México,2 Department of Animal Health and Biomedical Sciences and Genome Center of Wisconsin,3 Laboratory of Genetics, University of Wisconsin—Madison, Madison, Wisconsin 537064

Received 17 October 2001/ Returned for modification 5 January 2002/ Accepted 8 June 2002

The mechanisms underlying the adherence of Escherichia coli O157:H7 and other enterohemorrhagic E. coli (EHEC) strains to intestinal epithelial cells are poorly understood. We have identified a chromosomal region (designated lpfABCC'DE) in EHEC O157:H7 containing six putative open reading frames that was found to be closely related to the long polar (LP) fimbria operon (lpf) of Salmonella enterica serovar Typhimurium, both in gene order and in conservation of the deduced amino acid sequences. We show that lpfABCC'DE is organized as an operon and that its expression is induced during the exponential growth phase. The lpf genes from EHEC strain EDL933 were introduced into a nonfimbriated (Fim-) E. coli K-12 strain, and the transformed strain produced fimbriae as visualized by electron microscopy and adhered to tissue culture cells. Anti-LpfA antiserum recognized a ca. 16-kDa LpfA protein when expressed under regulation of the T7 promoter system. The antiserum also cross-reacted with the LP fimbriae in immunogold electron microscopy and Western blot experiments. Isogenic E. coli O157:H7 lpf mutants derived from strains 86-24 and AGT300 showed slight reductions in adherence to tissue culture cells and formed fewer microcolonies compared with their wild-type parent strains. The adherence and microcolony formation phenotypes were restored when the lpf operon was introduced on a plasmid. We propose that LP fimbriae participate in the interaction of E. coli O157:H7 with eukaryotic cells by assisting in microcolony formation.

* Corresponding author. Mailing address: Center for Vaccine Development and Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201. Phone: (410) 706-2344. Fax: (410) 706-0182. E-mail: jkaper{at}umaryland.edu.

Editor: A. D. O'Brien

Infection and Immunity, October 2002, p. 5416-5427, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5416-5427.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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