Identification and characterization of lpfABCC'DE, a fimbrial operon of enterohemorrhagic Escherichia coli O157

H7

Alfredo Torres, Jorge A. Giron, Nicole T. Perna, Valerie Burland, Fred R. Blattner, Fabiola Avelino-Flores, James B. Kaper

Research output: Contribution to journalArticle

149 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)5416-5427
Number of pages12
JournalInfection and Immunity
Volume70
Issue number10
DOIs
StatePublished - Oct 2002
Externally publishedYes

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Enterohemorrhagic Escherichia coli
Escherichia coli O157
Operon
Immune Sera
Electron Microscopy
Cell Culture Techniques
Salmonella enterica
Gene Order
Eukaryotic Cells
Open Reading Frames
Amino Acid Sequence
Plasmids
Western Blotting
Epithelial Cells
Escherichia coli
Phenotype
Growth
Genes
Proteins

ASJC Scopus subject areas

  • Immunology

Cite this

Identification and characterization of lpfABCC'DE, a fimbrial operon of enterohemorrhagic Escherichia coli O157 : H7. / Torres, Alfredo; Giron, Jorge A.; Perna, Nicole T.; Burland, Valerie; Blattner, Fred R.; Avelino-Flores, Fabiola; Kaper, James B.

In: Infection and Immunity, Vol. 70, No. 10, 10.2002, p. 5416-5427.

Research output: Contribution to journalArticle

Torres, Alfredo ; Giron, Jorge A. ; Perna, Nicole T. ; Burland, Valerie ; Blattner, Fred R. ; Avelino-Flores, Fabiola ; Kaper, James B. / Identification and characterization of lpfABCC'DE, a fimbrial operon of enterohemorrhagic Escherichia coli O157 : H7. In: Infection and Immunity. 2002 ; Vol. 70, No. 10. pp. 5416-5427.
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