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 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 in C. 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.
Pei
Z
Z
Department of Medicine, Vanderbilt University School of Medicine and Veterans Affairs Medical Center, Nashville, Tennessee 37232, USA.
Burucoa
C
C
Grignon
B
B
Baqar
S
S
Huang
X Z
XZ
Kopecko
D J
DJ
Bourgeois
A L
AL
Fauchere
J L
JL
Blaser
M J
MJ
D000818:Animals; D000942:Antigens, Bacterial; D001422:Bacterial Adhesion; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D016123:Campylobacter jejuni / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000032:analysis; D004847:Epithelial Cells / Q000382:microbiology; D005656:Fungal Proteins / Q000032:analysis; D006367:HeLa Cells; D006801:Humans; D007422:Intestines / Q000382:microbiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D018160:Receptors, Cytoplasmic and Nuclear / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins; D014169:Transformation, Bacterial
D000942:Antigens, Bacterial; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; C064042:CBF1 protein, S cerevisiae; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C489977:PEB1 protein, Campylobacter; D018160:Receptors, Cytoplasmic and Nuclear; D029701:Saccharomyces cerevisiae Proteins