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Identification and molecular profiling of a novel homolog of cystatin C from rock bream (Oplegnathus fasciatus) evidencing its transcriptional sensitivity to pathogen infections

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Abstract

Cystatins are reversible inhibitors of cysteine proteases which show an omnipresent distribution in the life on earth. Although, cystatins with mammalian origin were well characterized and their roles in physiology were reported in details, those from teleostean origin are still underrepresented in literature. However, role of cystatins in fish physiology and immune defense is highlighted in few recent reports. In this study, a cystatin C holmologue from rock bream (Oplegnathus fasciatus); termed RbCytC was identified and molecularly characterized. The complete coding sequence of RbCytC was 387 bp in length, which codes for a polypeptide with 129 amino acids, including a signal peptide of 19 amino acids. The consensus cystatin family signatures including a G residue, turning up towards the N-terminus region, QVVAG motif, locating at the middle of the sequence and the PW motif at the c terminal region was found to be well conserved in RbCytC. Phylogenetic analysis using different cystatin counterparts affirmed the close evolutionary relationship of RbCytC with its teleostan homologs which belong to family 2 cystatins. The predicted molecular model of RbCytC resembled most of the structural features of empirically elucidated tertiary structures for chicken egg white cystatin. According to the qPCR assays, RbCytC showed detectable expression in all fish tissues used in the experiment, with markedly pronounced expression level in liver. Moreover, its basal mRNA expression was up-regulated in liver and spleen tissues by experimental rock bream iridovirus infection, whereas down regulated in the same tissues, post live Edwardsiella tarda injection. Collectively, outcomes of our study validate the structural homology of RbCytC with known cystatin C similitudes, especially those of teleosts and suggest its potential roles in proteolytic processes of rock bream physiology as well as in host immune defense mechanisms.

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Acknowledgements

This research partially represents the project titled ‘Fish Vaccine Research Center’, funded by the Ministry of Oceans and Fisheries, Korea, and was supported by the Basic Science Research Program implemented by National Research Foundation of Korea with the funds provided by the Ministry of Education (2019R1A6A1A03033553).

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Authors and Affiliations

  1. Department of Basic Sciences and Social Science for Nursing, Faculty of Nursing, University of Colombo, Thalapathpitiya, 10250, Nugegoda, Sri Lanka

    Don Anushka Sandaruwan Elvitigala

  2. Department of Marine Life Sciences & Fish Vaccine Research Center, Jeju National University, 102 Jejudaehakno, Ara-Dong, Jeju, Jeju Self-Governing Province, 63243, Republic of Korea

    Jehee Lee

  3. Marine Science Institute, Jeju National University, Ara-Dong, Jeju,, Jeju Self-Governing Province, 63333, Republic of Korea

    Jehee Lee

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  1. Don Anushka Sandaruwan Elvitigala
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Contributions

Don Anushka Sandaruwan Elvitigala: Conceptualization, Data curation, Formal analysis, Investigation, Methodology, Writing- original draft. Jehee Lee: Conceptualization, Funding acquisition, Project administration, Writing- review and editing.

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Correspondence to Don Anushka Sandaruwan Elvitigala or Jehee Lee.

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Fish care and challenge experiments were reviewed and approved by Animal Care and Use Committee of Jeju National University.

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Elvitigala, D.A.S., Lee, J. Identification and molecular profiling of a novel homolog of cystatin C from rock bream (Oplegnathus fasciatus) evidencing its transcriptional sensitivity to pathogen infections. Mol Biol Rep 48, 4933–4942 (2021). https://doi.org/10.1007/s11033-021-06415-9

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