Identification, Expression Profiling, Microbial Binding, and Agglutination Analyses of Two Cathepsin B Genes in Black Rockfish (<i>Sebastes schlegelii</i>)

As a lysosomal cysteine protease of the papain subfamily, cathepsin B (CTSB) is characterized by its innate immune functions and hydrolytic activity. However, the functions of CTSB in the immune responses of teleosts remain to be clarified. In this study, two CTSB genes in <i>S. schlegelii<...

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Main Authors: Xinghua Zhuang, Xingchun Li, Wenpeng Li, Xuan Xu, Fengjun Lin, Yiying Liu, Chonghui Chen, Xiaoxu Zhang, Pei Zhang, Chao Li, Qiang Fu
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Marine Drugs
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Online Access:https://www.mdpi.com/1660-3397/23/5/213
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author Xinghua Zhuang
Xingchun Li
Wenpeng Li
Xuan Xu
Fengjun Lin
Yiying Liu
Chonghui Chen
Xiaoxu Zhang
Pei Zhang
Chao Li
Qiang Fu
author_facet Xinghua Zhuang
Xingchun Li
Wenpeng Li
Xuan Xu
Fengjun Lin
Yiying Liu
Chonghui Chen
Xiaoxu Zhang
Pei Zhang
Chao Li
Qiang Fu
author_sort Xinghua Zhuang
collection DOAJ
description As a lysosomal cysteine protease of the papain subfamily, cathepsin B (CTSB) is characterized by its innate immune functions and hydrolytic activity. However, the functions of CTSB in the immune responses of teleosts remain to be clarified. In this study, two CTSB genes in <i>S. schlegelii</i>, <i>Ss</i>CTSBa and <i>Ss</i>CTSBb, were identified. Both <i>Ss</i>CTSBa and <i>Ss</i>CTSBb are composed of a 993 bp ORF encoding 330 amino acids. It was found in a phylogeny analysis that both genes form monophyletic clades with their orthologous counterparts of Honeycomb rockfish (<i>Sebastes umbrosus</i>). A synteny analysis indicated that the CTSB homologues were comparatively conserved during vertebrate evolution. Additionally, quantitative real-time PCR revealed the ubiquitous mRNA expression of <i>Ss</i>CTSBa and <i>Ss</i>CTSBb in all of the examined tissues, and substantially differential expression patterns could be observed following <i>Aeromonas salmonicida</i> infection. A subcellular localization analysis demonstrated that the distribution of <i>Ss</i>CTSBa and <i>Ss</i>CTSBb was mainly in the cytoplasm. Moreover, r<i>Ss</i>CTSBa and r<i>Ss</i>CTSBb showed strong binding to Poly(I:C) and exhibited diverse agglutination effects on different bacteria. Overall, these findings suggest that the CTSB genes in black rockfish might show essential functions in the host defense of teleosts against bacterial infections, providing valuable insights for further investigations into the immune mechanism of teleost CTSB.
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spelling doaj-art-ce88e2fd76fd433286566d4a5e30c8182025-08-20T01:56:16ZengMDPI AGMarine Drugs1660-33972025-05-0123521310.3390/md23050213Identification, Expression Profiling, Microbial Binding, and Agglutination Analyses of Two Cathepsin B Genes in Black Rockfish (<i>Sebastes schlegelii</i>)Xinghua Zhuang0Xingchun Li1Wenpeng Li2Xuan Xu3Fengjun Lin4Yiying Liu5Chonghui Chen6Xiaoxu Zhang7Pei Zhang8Chao Li9Qiang Fu10School of Marine Science and Engineering, Qingdao Agricultural University, Qingdao 266109, ChinaSchool of Marine Science and Engineering, Qingdao Agricultural University, Qingdao 266109, ChinaHaidu College, Qingdao Agricultural University, Laiyang 265200, ChinaSchool of Marine Science and Engineering, Qingdao Agricultural University, Qingdao 266109, ChinaSchool of Marine Science and Engineering, Qingdao Agricultural University, Qingdao 266109, ChinaSchool of Marine Science and Engineering, Qingdao Agricultural University, Qingdao 266109, ChinaSchool of Marine Science and Engineering, Qingdao Agricultural University, Qingdao 266109, ChinaSchool of Marine Science and Engineering, Qingdao Agricultural University, Qingdao 266109, ChinaSchool of Marine Science and Engineering, Qingdao Agricultural University, Qingdao 266109, ChinaSchool of Marine Science and Engineering, Qingdao Agricultural University, Qingdao 266109, ChinaSchool of Marine Science and Engineering, Qingdao Agricultural University, Qingdao 266109, ChinaAs a lysosomal cysteine protease of the papain subfamily, cathepsin B (CTSB) is characterized by its innate immune functions and hydrolytic activity. However, the functions of CTSB in the immune responses of teleosts remain to be clarified. In this study, two CTSB genes in <i>S. schlegelii</i>, <i>Ss</i>CTSBa and <i>Ss</i>CTSBb, were identified. Both <i>Ss</i>CTSBa and <i>Ss</i>CTSBb are composed of a 993 bp ORF encoding 330 amino acids. It was found in a phylogeny analysis that both genes form monophyletic clades with their orthologous counterparts of Honeycomb rockfish (<i>Sebastes umbrosus</i>). A synteny analysis indicated that the CTSB homologues were comparatively conserved during vertebrate evolution. Additionally, quantitative real-time PCR revealed the ubiquitous mRNA expression of <i>Ss</i>CTSBa and <i>Ss</i>CTSBb in all of the examined tissues, and substantially differential expression patterns could be observed following <i>Aeromonas salmonicida</i> infection. A subcellular localization analysis demonstrated that the distribution of <i>Ss</i>CTSBa and <i>Ss</i>CTSBb was mainly in the cytoplasm. Moreover, r<i>Ss</i>CTSBa and r<i>Ss</i>CTSBb showed strong binding to Poly(I:C) and exhibited diverse agglutination effects on different bacteria. Overall, these findings suggest that the CTSB genes in black rockfish might show essential functions in the host defense of teleosts against bacterial infections, providing valuable insights for further investigations into the immune mechanism of teleost CTSB.https://www.mdpi.com/1660-3397/23/5/213cathepsinteleostexpression patterninnate immunity
spellingShingle Xinghua Zhuang
Xingchun Li
Wenpeng Li
Xuan Xu
Fengjun Lin
Yiying Liu
Chonghui Chen
Xiaoxu Zhang
Pei Zhang
Chao Li
Qiang Fu
Identification, Expression Profiling, Microbial Binding, and Agglutination Analyses of Two Cathepsin B Genes in Black Rockfish (<i>Sebastes schlegelii</i>)
Marine Drugs
cathepsin
teleost
expression pattern
innate immunity
title Identification, Expression Profiling, Microbial Binding, and Agglutination Analyses of Two Cathepsin B Genes in Black Rockfish (<i>Sebastes schlegelii</i>)
title_full Identification, Expression Profiling, Microbial Binding, and Agglutination Analyses of Two Cathepsin B Genes in Black Rockfish (<i>Sebastes schlegelii</i>)
title_fullStr Identification, Expression Profiling, Microbial Binding, and Agglutination Analyses of Two Cathepsin B Genes in Black Rockfish (<i>Sebastes schlegelii</i>)
title_full_unstemmed Identification, Expression Profiling, Microbial Binding, and Agglutination Analyses of Two Cathepsin B Genes in Black Rockfish (<i>Sebastes schlegelii</i>)
title_short Identification, Expression Profiling, Microbial Binding, and Agglutination Analyses of Two Cathepsin B Genes in Black Rockfish (<i>Sebastes schlegelii</i>)
title_sort identification expression profiling microbial binding and agglutination analyses of two cathepsin b genes in black rockfish i sebastes schlegelii i
topic cathepsin
teleost
expression pattern
innate immunity
url https://www.mdpi.com/1660-3397/23/5/213
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