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Functional Characterization and Transcriptional Analysis of clpP of Xanthomonas campestris pv. campestris

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Abstract

The caseinolytic protease (Clp) system is essential for survival under stress conditions and for virulence in several pathogenic bacteria. Xanthomonas campestris pv. campestris (Xcc) is a plant pathogen which causes black rot disease in crucifers. In this study, the Xcc clpP gene which is annotated to encode the proteolytic core of Clp was characterized. Mutation of clpP resulted in susceptibility to high temperature and puromycin stresses. Site-directed mutagenesis revealed that S105, H130, and D179 are critical amino acid residues for ClpP function in puromycin tolerance. Inactivation of clpP also revealed an attenuation of virulence on the host plant and a reduction in the production of extracellular cellulase, mannanase, pectinase, and protease. The affected phenotypes of the clpP mutant could be complemented to wild-type levels by the intact clpP gene. Transcriptional analysis revealed that expression of clpP is induced under heat shock condition.

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Acknowledgements

This study was supported by the Ministry of Science and Technology of Taiwan (Grant No. MOST107-2313-B-166-001-MY3).

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  1. Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Taichung, 406, Taiwan

    Chih-En Li, Chao-Tsai Liao, Hsueh-Hsia Lo & Yi-Min Hsiao

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  1. Chih-En Li
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Contributions

CE carried out the experiments and prepared figures and tables. CE, CT, HH, and YM analyzed the experimental results and interpreted the data. YM managed the grants, supervised the laboratory work, led the design, and coordination of this study and wrote the manuscript draft. All authors read and approved the final manuscript.

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Correspondence to Yi-Min Hsiao.

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Li, CE., Liao, CT., Lo, HH. et al. Functional Characterization and Transcriptional Analysis of clpP of Xanthomonas campestris pv. campestris. Curr Microbiol 77, 2876–2885 (2020). https://doi.org/10.1007/s00284-020-02093-1

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