Diffuse adherence of enteropathogenic Escherichia coli strains--processing of AIDA-I

The adherence of pathogenic Escherichia coli to the mucosa of the small intestine is an important step in the development of diarrhoea. To study the molecular basis of the diffuse adherence (DA) pattern of E. coli strains expressing the classical serotypes of enteropathogenic E. coli (EPEC), strain 2787 (O126:H27) was investigated. By expression cloning, a plasmid-derived 6.0 kb DNA fragment was identified which conferred the DA phenotype on recipient K-12 strains. This fragment encoded the 100 kDa adhesin involved in diffuse adherence (AIDA-I) which by mild heat shock treatment was isolated from the surface of the wild-type and recombinant DA-positive strains. Analysis of the entire DNA fragment revealed two open reading frames coding for proteins of 45 kDa and 132 kDa, respectively. The 132 kDa protein has been identified as the AIDA-I precursor protein which after cleavage of the signal sequence undergoes additional C-terminal processing for maturation to AIDA-I. Though the function of the cytoplasmic 45 kDa protein is not known, preliminary evidence indicates that authentic expression of the protein is a prerequisite for the correct processing of the 132 kDa precursor to AIDA-I. The AIDA-I precursor exhibits significant homology to the virG (icsA) protein of Shigella flexneri which apparently plays a major role in the events leading to the intercellular spread of invasive Shigella organisms.