Mutation in the peb1A Locus of Campylobacter jejuni Reduces Interactions with Epithelial Cells and Intestinal Colonization of Mice

Abstract

Campylobacter jejuni is one of the leading causes of bacterial diarrhea throughout the world. We previously found that PEB1 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, contains 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 inC. jejuni adherence, peb1A, the gene encoding PEB1, was disrupted in strain 81-176 by insertion of a kanamycin resistance gene through homologous recombination. Inactivation of this operon completely abolished expression of CBF1, as determined by sodium dodecyl sulfate-polyacrylamide 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 invasion of epithelial cells in culture. Mouse challenge studies showed that the rate and duration of intestinal colonization by the mutant were significantly lower and shorter than with the wild-type strain. In summary, PEB1 is identical to a previously identified cell-binding factor, CBF1, in C. jejuni, and the peb1A locus plays an important role in epithelial cell interactions and in intestinal colonization in a mouse model.

The Campylobacter jejuni PEB1a adhesin is an aspartate/glutamate‐binding protein of an ABC transporter essential for microaerobic growth on dicarboxylic amino acids

“…From the results reported here, it is clear that in addition to its well characterized role as an adhesin, the transport function of PEB1a and the PEB1 system in toto is key to the ability of C. jejuni to catabolize aspartate and glutamate, two common amino acids likely to be present in gut contents. This suggests a re-interpretation of the in vivo significance of PEB1a, as the colonization defect previously reported for a peb1a mutant in the mouse model of infection (Pei et al, 1998) could actually be due to one or both of these functions. In order to assess the in vivo contribution of adhesion versus transport, it would be informative to study the colonization potential of a strain which produced an engineered PEB1a unable to bind its ligands but otherwise wild type for localization and adhesion.…”

An ATP‐binding cassette‐type cysteine transporter in Campylobacter jejuni inferred from the structure of an extracytoplasmic solute receptor protein

“…The role of Cj0982 on the outside of the cell is unknown. Interestingly, two other C. jejuni proteins that are expected to have ESR-like folds, Peb1 and Peb3, are crucial for adhesion and virulence (Pei et al, 1998).…”

Relaxation of DNA supercoiling leads to increased invasion of epithelial cells and protein secretion by Campylobacter jejuni

“…A number of C. jejuni virulence factors have been shown to contribute to C. jejuni adhesion and invasion in-vitro ( O Cr oin ın and Backert, 2012). It is clear that production of Peb1 and Peb4 are important for the ability of C. jejuni to adhere to the epithelial cell boundary but their influence on C. jejuni adhesion/invasion appears to be indirect (Pei et al, 1991;Kervella et al, 1993;Burucoa et al, 1995;Pei et al, 1998;Kale et al, 2011). JlpA is a 42 kDa C. jejuni specific adhesin, which is surface exposed.…”