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Infection and Immunity, May 1999, p. 2292-2298, Vol. 67, No. 5
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Characterization of the Escherichia coli AF/R1 Pilus Operon: Novel Genes Necessary for Transcriptional Regulation and for Pilus-Mediated Adherence

J. Robert Cantey,1,* R. K. Blake,1 J. R. Williford,2 and Steve L. Moseley2

Ralph H. Johnson V. A. Medical Center and Infectious Diseases Division, Medical University of South Carolina, Charleston, South Carolina 29425,1 and Department of Microbiology, School of Medicine, University of Washington, Seattle, Washington 981952

Received 26 October 1998/Returned for modification 3 December 1998/Accepted 24 February 1999

We isolated the genetic determinant of AF/R1 pilus production in attaching/effacing Escherichia coli RDEC-1 and identified seven genes required for pilus expression and function. DNA sequence analysis of the structural subunit gene afrA corrected an error in the published sequence and extended homology with the F18 pilus subunit of pig edema E. coli strains. AfrB and AfrC, encoded downstream from AfrA, were required for pilus expression. AfrB was related to the usher protein PefC of Salmonella typhimurium plasmid-encoded fimbriae, and AfrC was related to PefD, a chaperone protein. AfrD and AfrE, encoded downstream from AfrC, were not necessary for the expression of AF/R1 pili but were required for ileal adherence as assayed by ileal brush border aggregation. Thus, the adhesive subunit of the AF/R1 pilus is distinct from the structural subunit, as is the case for Pap pili and type 1 pili. AfrD was related to FedE of the F18 fimbrial operon of the E. coli strain that causes edema disease in pigs. AfrE was a novel protein. AfrR and AfrS are encoded upstream from AfrA, in the opposite orientation. AfrR is related to the AraC family of transcriptional regulators, and AfrR and AfrS interact to function in a novel mode of transcriptional activation of afrA. AF/R1 pili mediate the adherence to Peyer's patch M cells, ileal mucosa, and colonic mucosa in a rabbit model of diarrhea caused by enteropathogenic E. coli. Our observations will facilitate the further study of the phenomena of M-cell adherence.

* Corresponding author. Mailing address: Infectious Diseases Division, Medical University of South Carolina, 100 Doughty St., P.O. Box 250752, Charleston, SC 29425. Phone: (803) 792-4541. Fax: (803) 792-6680. E-mail: canteyjr{at}musc.edu.

Infection and Immunity, May 1999, p. 2292-2298, Vol. 67, No. 5
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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