MUTATION IN THE PEB1A LOCUS OF CAMPYLOBACTER-JEJUNI REDUCES INTERACTIONS WITH EPITHELIAL-CELLS AND INTESTINAL COLONIZATION OF MICE

Campylobacter jejuni is one of the leading causes of bacterial diarrhe a throughout the world. We previously found that PERI is a homolog of cluster 3 binding proteins of bacterial ABC transporters and that a C. jejuni adhesin, cell-binding factor 1 (CBF1), if not identical to, co ntains PEB1. A single protein migrating at approximately 27 to 28 kDa was recognized by anti-CBF1 and anti-PEB1. To determine the role that the operon encoding PEB1 plays in C. jejuni adherence, peb1A, the gene encoding PEB1, was disrupted in strain 81-176 by insertion of a kanam ycin resistance gene through homologous recombination. Inactivation of this operon completely abolished expression of CBF1, as determined by sodium dodecyl sulfate-polyaccrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. In comparison to the wild-type strain, the mutant strain showed 50- to 100-fold less adherence to and 15-fold less inva sion of epithelial cells in culture, Mouse challenge studies showed th at the rate and duration of intestinal colonization by the mutant were significantly lower and shorter than with the wild-type strain. In su mmary, PEB1 is identical to a previously identified cell-binding facto r, CBF1, in C. jejuni, and the peb1A locus plays an important role in epithelial cell interactions and in intestinal colonization in a mouse model.