Complete nucleotide sequence and analysis of the locus of enterocyte effacement from rabbit diarrheagenic Escherichia coli RDEC-1

The pathogenicity island termed the locus of enterocyte effacement (LEE) is found in diverse attaching and effacing pathogens associated with diarrhea in humans and other animal species. To explore the relation of variation i n LEE sequences to host specificity and genetic lineage, we determined the nucleotide sequence of the LEE region from a rabbit diarrheagenic Escherich ia coli strain RDEC-1 (O15:H-) and compared it with those from human entero pathogenic E. coli (EPEC, O127:H6) and enterohemorrhagic E. coli (EHEC, O15 7:H7) strains. Differing from EPEC and EHEC LEEs, the RDEC-1 LEE is not ins erted at selC and is flanked by an IS2 element and the lifA toxin gene. The RDEC-1 LEE contains a core region of 40 open reading frames, all of which are shared with the LEE of EPEC and EHEC. orf3 and the ERIC (enteric repeti tive intergenic consensus) sequence present in the LEEs of EHEC and EPEC ar e absent from the RDEC-1 LEE. The predicted promoters of LEE1, LEE2, LEE3, tir, and LEE4 operons are highly conserved among the LEEs, although the ups tream regions varied considerably for tir and the crucial LEE2 promoter, su ggesting differences in regulation. Among the shared genes, high homology ( >95% identity) between the RDEC-1 and the EPEC and EHEC LEEs at the predict ed amino acid level was observed for the components of the type LII secreti on apparatus, the Ces chaperones, and the Ler regulator. In contrast, more divergence (66 to 88% identity) was observed in genes encoding proteins inv olved in host interaction, such as intimin (Eae) and the secreted proteins (Tir and Esps). A comparison of the highly variable genes from RDEC-1 with those from a number of attaching and effacing pathogens infecting different species and of different evolutionary lineages was performed. Although RDE C-1 diverges from some human-infecting EPEC and EHEC, most of the variation observed appeared to be due to evolutionary lineage rather than host speci ficity. Therefore, much of the observed hypervariability in genes involved in pathogenesis may not represent specific adaptation to different host spe cies.