title: Identification and characterization of lpfABCC'DE, a fimbrial operon of enterohemorrhagic Escherichia coli O157:H7.
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.
authors.affiliation: Center for Vaccine Development and Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA.
authors.forename: Alfredo G
authors.initials: AG
authors.lastname: Torres
authors.forename: Jorge A
authors.initials: JA
authors.lastname: Giron
authors.forename: Nicole T
authors.initials: NT
authors.lastname: Perna
authors.forename: Valerie
authors.initials: V
authors.lastname: Burland
authors.forename: Fred R
authors.initials: FR
authors.lastname: Blattner
authors.forename: Fabiola
authors.initials: F
authors.lastname: Avelino-Flores
authors.forename: James B
authors.initials: JB
authors.lastname: Kaper
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial; D001422:Bacterial Adhesion / Q000235:genetics / Q000502:physiology; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D003429:Cross Reactions; D004269:DNA, Bacterial / Q000235:genetics; D004751:Enteritis / Q000209:etiology / Q000382:microbiology; D004927:Escherichia coli Infections / Q000209:etiology / Q000382:microbiology; D019453:Escherichia coli O157 / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology / Q000648:ultrastructure; D010861:Fimbriae, Bacterial / Q000235:genetics* / Q000276:immunology / Q000502:physiology / Q000648:ultrastructure; D015870:Gene Expression; D005798:Genes, Bacterial; D006367:HeLa Cells; D006801:Humans; D008854:Microscopy, Electron; D009876:Operon
chemical_list: D000907:Antibodies, Bacterial; D004269:DNA, Bacterial