The Escherichia coli ycbQRST operon encodes fimbriae with laminin-binding and epithelial cell adherence properties in Shiga-toxigenic E.  coli O157:H7.
The human pathogen Shiga-toxigenic Escherichia coli (STEC) O157:H7 contains a ycbQRST fimbrial-like operon, which shares significant homology to the family of F17 fimbrial biogenesis genes f17ADCG found in enterotoxigenic E.  coli. We report that growth of STEC O157:H7 strain EDL933 in minimal Minca medium at 37°C and during adherence to epithelial cells led to the production of fine peritrichous fimbriae, which were found to be composed of a major subunit of 18  kDa whose N-terminal amino acid sequence matched the predicted protein product of the ycbQ gene; and showed significant homology to the F17a-A fimbrin. Similar to the F17 fimbriae, the purified STEC fimbriae and the recombinant YcbQ protein fused to a His peptide tag bound laminin, but not fibronectin or collagen. Thus, we propose the name E.  coli YcbQ laminin-binding fimbriae (ELF) to designate the fimbriae encoded by the ycbQRST operon. The role of ELF as an adherence factor of STEC to cultured epithelial cells was investigated. We provide compelling evidence demonstrating that ELF contributes to adherence of STEC to human intestinal epithelial cells and to cow and pig gut tissue in vitro. Deletion in the fimbrin subunit gene elfA (or ycbQ) in STEC strain EDL933 led to an isogenic strain, which showed significant reduction (60%) in adherence to HEp-2 cells in comparison with the parental strain. In addition, antibodies against the purified ELF also partially blocked adherence of two STEC O157:H7 strains. These observations suggest that ELF functions as an accessory adherence factor that, along with other known redundant adhesins, contributes to the overall adhesive properties of STEC O157:H7 providing these organisms with ecological advantages to survive in different hosts and in the environment.
Samadder
Partha
P
Department of Immunobiology, University of Arizona, Tucson, AZ  85724, USA.
Xicohtencatl-Cortes
Juan
J
Saldaña
Zeus
Z
Jordan
Dianna
D
Tarr
Phillip I
PI
Kaper
James B
JB
Girón
Jorge A
JA
D000595:Amino Acid Sequence; D000818:Animals; D001422:Bacterial Adhesion; D002417:Cattle; D002460:Cell Line; D004847:Epithelial Cells / Q000382:microbiology*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D036781:Fimbriae Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007797:Laminin / Q000378:metabolism*; D008969:Molecular Sequence Data; D009876:Operon; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid; D054323:Shiga-Toxigenic Escherichia coli / Q000472:pathogenicity*; D013552:Swine
D029968:Escherichia coli Proteins; D007797:Laminin; D036781:Fimbriae Proteins