Abstract
Vibrio alginolyticus is an opportunistic pathogen in both humans and marine animals. Collagenase encoded by colA is considered to be one of the virulence factors. Expression of colA is regulated by multiple environmental factors, e.g., temperature, growth phase, and substrate. To elucidate the mechanism of regulation of colA expression, transposon mutagenesis was performed. VarS, a sensor histidine kinase of the two-component regulatory system, was demonstrated to regulate the expression of colA. VarA, a cognate response regulator of VarS, was also identified and shown to be involved in the regulation of colA expression. In vitro phosphorylation assays showed that phosphorylated VarS acted as a phosphoryl group donor to VarA. A site-directed mutagenesis study showed that the His300, Asp718 and His874 residues in VarS were essential for the phosphorylation of VarS, and the Asp54 residue in VarA was likely to receive the phosphoryl group from VarS. The results demonstrate that the VarS/VarA two-component regulatory system regulates the expression of collagenase in V. alginolyticus.
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Acknowledgements
The mariner-based transposon delivery plasmid, pSC189, was kindly provided by Dr. John Mekalanos (Department of Microbiology and Immunobiology, Harvard Medical School, Boston, USA). RHO3, DH5α(λpir), pEX18Ap and pUC18T-mini-Tn7-Gm were kindly provided by Dr. Herbert P. Schweizer (Department of Molecular Genetics and Microbiology, Emerging Pathogens Institute, University of Florida, Florida, USA). This work was financially supported by JSPS KAKENHI Grant Number 26860290 and the Takeda Science Foundation.
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Mima, T., Gotoh, K., Yamamoto, Y. et al. Expression of Collagenase is Regulated by the VarS/VarA Two-Component Regulatory System in Vibrio alginolyticus . J Membrane Biol 251, 51–63 (2018). https://doi.org/10.1007/s00232-017-9991-9
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DOI: https://doi.org/10.1007/s00232-017-9991-9