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Two Distinct C-Type Lysozymes in Goldfish: Molecular Characterization, Antimicrobial Potential, and Transcriptional Regulation in Response to Opposing Effects of Bacteria/Lipopolysaccharide and Dexamethasone/Leptin

by Ting Chen 1,2,3,†, Yingzhu Rao 1,†, Jiaxi Li 4, Chunhua Ren 2,3, Dongsheng Tang 4, Tiehao Lin 5, Jiatai Ji 3,6, Rong Chen 1,* and Aifen Yan 4,*
1
Institute of Applied Biotechnology, School of Life Science and Technology, Lingnan Normal University, Zhanjiang 528225, China
2
CAS Key Laboratory of Tropical Marine Bio-Resources and Ecology (LMB), Guangdong Provincial Key Laboratory of Applied Marine Biology (LAMB), South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China
3
Institution of South China Sea Ecology and Environmental Engineering (ISEE), Chinese Academy of Sciences, Guangzhou 510301, China
4
School of Stomatology and Medicine, School of Life Science and Engineering, Foshan University, Foshan 528000, China
5
Microbiological department, Guangdong Institute for Drug Control, Guangzhou 510663, China
6
Guangdong Haimao Investment Co., Ltd., Zhanjiang 524001, China
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Int. J. Mol. Sci. 2020, 21(2), 501; https://doi.org/10.3390/ijms21020501
Received: 15 December 2019 / Revised: 8 January 2020 / Accepted: 10 January 2020 / Published: 13 January 2020
Lysozymes are key antimicrobial peptides in the host innate immune system that protect against pathogen infection. In this study, the full-length cDNAs of two c-type lysozymes (gfLyz-C1 and gfLyz-C2) were cloned from goldfish (Carassius auratus). The structural domains, three-dimensional structures, and amino acid sequences of gfLyz-C1 and gfLyz-C2 were highly comparable, as the two proteins shared 89.7% sequence identity. The gfLyz-C1 and gfLyz-C2 recombinant proteins were generated in the insoluble fractions of an Escherichia coli system. Based on the results of lysoplate and turbidimetric assays, gfLyz-C1 and gfLyz-C2 showed broad-spectrum antimicrobial properties with high levels of activity against Micrococcus lysodeikticus, Vibrio parahemolyticus, and Edwardsiella tarda, and relatively low activity against E. coli. Both gfLyz-C1 and gfLyz-C2 mRNAs were mainly expressed in the trunk kidney and head kidney, and gfLyz-C1 was expressed at much higher levels than gfLyz-C2 in the corresponding tissues. The expression of the gfLyz-C1 and gfLyz-C2 transcripts in the trunk kidney and head kidney was induced in these tissues by challenge with heat-inactivated E. coli and lipopolysaccharides (LPS), and the transcriptional responses of gfLyz-C1 were more intense. In goldfish primary trunk kidney cells, the levels of the gfLyz-C1 and gfLyz-C2 transcripts were upregulated by heat-inactivated E. coli, V. parahemolyticus, and E. tarda, as well as LPS, and downregulated by treatment with dexamethasone and leptins. Overall, this study may provide new insights that will improve our understanding of the roles of c-type lysozymes in the innate immunity of cyprinid fish, including the structural and phylogenetic characteristics, antimicrobial effects, and regulatory mechanism. View Full-Text
Keywords: c-type lysozyme; fish; antimicrobial activity; innate immunity; dexamethasone; leptin c-type lysozyme; fish; antimicrobial activity; innate immunity; dexamethasone; leptin
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MDPI and ACS Style

Chen, T.; Rao, Y.; Li, J.; Ren, C.; Tang, D.; Lin, T.; Ji, J.; Chen, R.; Yan, A. Two Distinct C-Type Lysozymes in Goldfish: Molecular Characterization, Antimicrobial Potential, and Transcriptional Regulation in Response to Opposing Effects of Bacteria/Lipopolysaccharide and Dexamethasone/Leptin. Int. J. Mol. Sci. 2020, 21, 501.

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