Functional Analysis of the Campylobacter jejuni cj0183 and cj0588 Genes

Abstract

The cj0183 and cj0588 genes identified in the Campylobacter jejuni NCTC 11168 genome encode proteins with amino acid sequences predicted to be homologous to other bacterial hemolysins. The Cj0183 protein exhibits homology to Brachyspira hyodysenteriae TlyC protein, whereas the cj0588 gene product is homologous to TlyA proteins Brachyspira hyodysenteriae, Helicobacter pylori, and Mycobacterium tuberculosis, which play a crucial role in bacterial virulence. The aim of our work was to examine the hemolytic activity and determine the role of cj0183- and cj0588-encoded proteins on the adherence of chosen C. jejuni strains to the Caco-2 cell line by constructing deletion mutants in the mentioned genes. We found out there is no difference in hemolytic activity between both mutants in gene cj0183 and cj0588 and the wild strains. However, Cj0588 protein but not Cj0183 is involved in adherence to the Caco-2 cells.

Analysis of Campylobacter jejuni Whole-Genome DNA Microarrays: Significance of Prophage and Hypervariable Regions for Discriminating Isolates

“…Two of these, the hemolysin gene tylA (Cj0588) and the heme transport gene chuA (Cj1614), were NCTC11168 genes. The tylA gene is involved in cell adherence and appears to have an important role in host colonization and virulence (Salamaszyń ska-Guz and Klimuszko, 2008). The divergence of tylA within populations of C. jejuni has not been well characterized, but its role in virulence warrants further investigation as a genotyping marker.…”

Genotyping and PCR detection of potential virulence genes in Campylobacter jejuni and Campylobacter coli isolates from different sources in Poland

“…They have detected the presence of antiCj0588 antibodies in the blood serum of patients after C. jejuni infection. The cj0588 gene was determined to encode a protein involved in adherence to Caco-2 cells (Sałamaszyńska-Guz and Klimuszko 2008).…”

Influence of mutation in cj0183 and cj0588 genes for colonization abilities of Campylobacter jejuni in Caco-2 cells using confocal laser scanning microscope

“…Adhesion and invasion assays for cj0183 and cj0588 mutants were performed as described by Sałamaszyńska-Guz and Klimuszko (2008). The C. jejuni cells and the Caco-2 cell tubulin were visualized using mouse monoclonal anti-bovine -tubulin antibodies, Alexa Fluor 546 conjugated goat anti-mouse IgG (Molecular Probes Inc.) and polyclonal anti -Campylobacter jejuni -FITC conjugate (Fitzgerald).…”

“…This mutation reduces both adhesion and internalization of 81-176 and 81116 C. jejuni strains to the epithelial cell line (Sałamaszyńska-Guz and Klimuszko 2008 (Hyatt et al 1994;Martino et al 2001). The aim of the presented work was to determine in vitro adherence and an invasion of Caco-2 cells by constructed mutants in both cj0183 and cj0588 genes in 81116 and 81-176 strain (81116-ΔCj0588Cj0183 and 81-176-ΔCj0588Cj0183).…”