Campylobacter protein oxidation influences epithelial cell invasion or intracellular survival as well as intestinal tract colonization in chickens.
The Dsb family of redox proteins catalyzes disulfide bond formation and isomerization. Since mutations in dsb genes change the conformation and stability of many extracytoplasmic proteins, and since many virulence factors of pathogenic bacteria are extracytoplasmic, inactivation of dsb genes often results in pathogen attenuation. This study investigated the role of 2 membrane-bound oxidoreductases, DsbB and DsbI, in the Campylobacter jejuni oxidative Dsb pathway. Campylobacter mutants, lacking DsbB or DsbI or both, were constructed by allelic replacement and used in the human intestinal epithelial T84 cell line for the gentamicin protection assay (invasion assay) and chicken colonization experiments. In C. coli strain 23/1, the inactivation of the dsbB or dsbI gene separately did not significantly affect the colonization process. However, simultaneous disruption of both membrane-bound oxidoreductase genes significantly decreased the strain’s ability to colonize chicken intestines. Moreover, C. jejuni strain 81-176 with mutated dsbB or dsbI genes showed reduced invasion/intracellular survival abilities. No cells of the double mutants (dsbB⁻ dsbI⁻) of C. jejuni 81-176 were recovered from human cells after 3 h of invasion.
Lasica
A M
AM
Department of Bacterial Genetics, Institute of Microbiology, Biology Faculty, University of Warsaw, Warsaw, Poland.
Wyszynska
A
A
Szymanek
K
K
Majewski
P
P
Jagusztyn-Krynicka
E K
EK
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D016123:Campylobacter jejuni / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D002460:Cell Line; D002645:Chickens / Q000382:microbiology*; D015169:Colony Count, Microbial; D019295:Computational Biology; D004847:Epithelial Cells / Q000166:cytology / Q000382:microbiology*; D041981:Gastrointestinal Tract / Q000382:microbiology*; D020868:Gene Silencing; D005798:Genes, Bacterial / Q000235:genetics; D006801:Humans; D042541:Intracellular Space / Q000382:microbiology*; D050296:Microbial Viability; D009154:Mutation / Q000235:genetics; D010084:Oxidation-Reduction; D020543:Proteome / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics
D001426:Bacterial Proteins; D020543:Proteome