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Infection and Immunity, August 2003, p. 4516-4525, Vol. 71, No. 8
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.8.4516-4525.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Characterization of the Novel Factor Paa Involved in the Early Steps of the Adhesion Mechanism of Attaching and Effacing Escherichia coli

Isabelle Batisson,1,2 Marie-Pierre Guimond,2 Francis Girard,2 Hongyan An,3 Chengru Zhu,4 Eric Oswald,5 John M. Fairbrother,2 Mario Jacques,2 and Josée Harel2*

Laboratoire de Biologie des Protistes, UMR 6023, Université Blaise Pascal, Aubière,1 Unité INRA-ENVT de Microbiologie Moléculaire, École Vétérinaire de Toulouse, 31076 Toulouse Cedex, France,5 Groupe de Recherche sur les Maladies Infectieuses du Porc, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec J2S 7C6, Canada,2 Advantage International USA, Inc., Westport, Connecticut 06880,3 Center for Vaccine Development, Division of Geographic Medicine, University of Maryland School of Medicine, Baltimore, Maryland 212014

Received 3 December 2002/ Returned for modification 10 February 2003/ Accepted 20 May 2003

Nonenterotoxigenic porcine Escherichia coli strains belonging to the serogroup O45 have been associated with postweaning diarrhea in swine and adhere to intestinal epithelial cells in a characteristic attaching and effacing (A/E) pattern. O45 porcine enteropathogenic E. coli (PEPEC) strain 86-1390 induces typical A/E lesions in a pig ileal explant model. Using TnphoA transposon insertion mutagenesis on strain 86-1390, we found a mutant that did not induce A/E lesions. The insertion was identified in a gene designated paa (porcine A/E-associated gene). Sequence analysis of paa revealed an open reading frame of 753 bp encoding a 27.6-kDa protein which displayed 100, 51.8, and 49% homology with Paa of enterohemorrhagic E. coli O157:H7 strains (EDL933 and Sakai), PEB3 of Campylobacter jejuni, and AcfC of Vibrio cholerae, respectively. Chromosomal localization studies indicated that the region containing paa was inserted between the yciD and yciE genes at about 28.3 min of the E. coli K-12 chromosome. The presence of paa and eae sequences in the porcine O45 strains is highly correlated with the A/E phenotype. However, the observation that three eae-positive but paa-negative PEPEC O45 strains were A/E negative provides further evidence for the importance of the paa gene in the A/E activity of O45 strains. As well, the complementation of the paa mutant restored the A/E activity of the 86-1390 strain, showing the involvement of Paa in PEPEC pathogenicity. These observations suggest that Paa contributes to the early stages of A/E E. coli virulence.

* Corresponding author. Mailing address: Groupe de Recherche sur les Maladies Infectieuses du Porc, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec J2S 7C6, Canada. Phone: (450) 773-8521, ext. 8233. Fax: (450) 778-8108. E-mail: josee.harel{at}umontreal.ca.

Editor: B. B. Finlay

Infection and Immunity, August 2003, p. 4516-4525, Vol. 71, No. 8
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.8.4516-4525.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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