Abstract
To identify virulence-associated genes of a fish pathogen Yersinia ruckeri, we screened a total of 1056 mini-Tn5-Km2 signature-tagged mutants in rainbow trout by immersion challenge. Of 1056, 25 mutants were found survival-defective as they could not be re-isolated from fish kidney 7 days after infection. Mutated gene in F2-4 mutant, one of the 25 mutants, was homologous to uvrY that encodes UvrY response regulator of BarA–UvrY two-component system (TCS). Mutant F2-4 was significantly more sensitive (P < 0.05) to H2O2-mediated killing and was less able to infect Epithelioma papulosum cyprini cells. However, UvrY mutation did not affect survival of F2-4 mutant in the presence of non-immune fish serum and its ability to grow under iron starvation. In a time-course co-infection, mutant F2-4 had lower bacterial loads on day 1 itself, and by day 5 there was nearly a 1,000-fold difference in infection levels of the parent and mutant strains. The barA homolog of Y. ruckeri was PCR-amplified and sequence analyses identified four domains that were characteristic of hybrid histidine kinases. To conclude, the BarA–UvrY TCS contributes to the pathogenesis of Y. ruckeri in its natural host rainbow trout, possibly by regulating invasion of epithelial cells and sensitivity to oxidative stress induced by immune cells.
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Acknowledgments
Financial support for this study was provided by the Natural Sciences and Engineering Research Council of Canada. The pUTmini-Tn5-Km2 signature tags were kindly provided by Dr. R. Levesque, University of Laval, Québec, Canada. Authors thank Steve Lord for providing the EPC cells and other reagents for cell invasion assays.
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Communicated by Jorge Membrillo-Hernández.
Accession numbers for DNA sequences: EU170243 (UvrY) and EU170244 (BarA).
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Dahiya, I., Stevenson, R.M.W. The UvrY response regulator of the BarA–UvrY two-component system contributes to Yersinia ruckeri infection of rainbow trout (Oncorhynchus mykiss). Arch Microbiol 192, 541–547 (2010). https://doi.org/10.1007/s00203-010-0582-8
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DOI: https://doi.org/10.1007/s00203-010-0582-8