title: Molecular evolution of the intimin gene in O111 clones of pathogenic Escherichia coli.
abstract: Intimin is an important virulence factor in two groups of enteric pathogens: enteropathogenic Escherichia coli (EPEC), which is a major cause of infant diarrhea in the developing world, and enterohemorrhagic E. coli (EHEC), which has caused large food-borne outbreaks of hemorrhagic colitis in the United States and other developed countries. Intimin is encoded on a 35-kb pathogenicity island called the locus of enterocyte effacement (LEE). At least five antigenic types have been described for the highly variable gene, and each type is generally characteristic of particular evolutionary lineages. We determined the nucleotide sequences of intimin and other LEE genes in two O111 clones that have not been amenable to typing. The sequences from both O111:H8 and O111:H9 differed from the Int-beta that is typical of other clones in the same evolutionary lineage. The sequence from the O111:H8 strains was a mosaic of divergent segments that alternately clustered with Int-alpha, Int-beta, or Int-gamma. The sequence from the O111:H9 clone consistently showed a close relationship with that from E2348/69, a distantly related strain that expresses Int-alpha. The results suggest that there have been multiple acquisitions of the LEE in the EHEC 2/EPEC 2 clonal lineage, with a recent turnover in either O111:H8 or its close relatives. Amino acid substitutions that alter residue charge occurred more frequently than would be expected under random substitution in the extracellular domains of intimin, suggesting that diversifying selection has promoted divergence in this region of the protein. An N-terminal domain that presumably functions in the periplasm may also be under positive selection.
authors.affiliation: Microbial Evolution Laboratory, National Food Safety and Toxicology Center, Michigan State University, East Lansing, Michigan 48824, USA.
authors.forename: Cheryl L
authors.initials: CL
authors.lastname: Tarr
authors.forename: Thomas S
authors.initials: TS
authors.lastname: Whittam
mesh_terms: D018829:Adhesins, Bacterial / Q000145:classification / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002352:Carrier Proteins / Q000145:classification / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000145:classification / Q000235:genetics / Q000472:pathogenicity; D019453:Escherichia coli O157 / Q000145:classification / Q000235:genetics* / Q000472:pathogenicity; D029968:Escherichia coli Proteins; D019143:Evolution, Molecular; D005798:Genes, Bacterial; D018740:Genetic Heterogeneity; D006801:Humans; D008969:Molecular Sequence Data; D009030:Mosaicism; D010802:Phylogeny; D011110:Polymorphism, Genetic; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
chemical_list: D018829:Adhesins, Bacterial; D002352:Carrier Proteins; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; C073819:eaeA protein, E coli