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大口黑鲈(Micropterus salmoides)酪氨酸蛋白激酶家族成员的分子特征、表达、进化分析及分子对接

Molecular characterization, expression, evolutionary analysis and molecular docking of the Janus activated kinase family members of largemouth bass (Micropterus salmoides).

作者信息

Pi Xiangyu, Pan Mingzhu, Zhang Qihuan, Wang Zisheng, Qi Zhitao

机构信息

School of Marine and Bioengineering, Yancheng Institute of Technology, Yancheng, China.

School of Marine and Bioengineering, Yancheng Institute of Technology, Yancheng, China; Jiangsu Key Laboratory for Exploration and Utilization of Marine Wetland Biological Resources, Yancheng Institute of Technology, Yancheng, China.

出版信息

Dev Comp Immunol. 2025 May;167:105375. doi: 10.1016/j.dci.2025.105375. Epub 2025 Apr 26.

Abstract

Janus kinases (JAKs) play crucial roles in the immune responses by binding the cytokine receptors. In the present study, five JAKs family members (JAK1, JAK2a, JAK2b, JAK3, and TYK2) were identified in largemouth bass (Micropterus salmoides). The five JAKs members of largemouth bass contained several conserved protein domains, including a FERM domain, an SH2 domain, a pseudokinase (STyrkc) domain, and a tyrosine kinase (Tyrkc) domain. Phylogenetic analysis revealed the JAK2/2a/2b and JAK3 formed a clade and the JAK1/TYK2 formed another separate clade. Realtime qPCR detection showed that all five JAKs genes were constitutively expressed in ten selected tissues, with highly expression in spleen and head kidney (HK). Following Edwardsiella piscicida infection, the five JAKs genes were significantly upregulated in the spleen and primary hepatocytes at different times post infection. Further selection pressure analysis revealed the five JAKs members underwent negative selection pressures during evolution. The FERM and Tyrkc domains of the five JAK members occurred purifying selection, and involved in interaction with STAT3, which confirmed by protein-protein interaction (PPI) network and molecular docking analysis. Also, molecular docking results indicated that the hydrogens bonds and salt bonds had crucial roles in the JAK/STAT3 complexes formation. Our results indicated that JAKs had important roles in the immune response against bacterial in largemouth bass, providing basis for elucidating the mechanism of JAK/STAT signaling pathway in fish.

摘要

Janus激酶(JAKs)通过结合细胞因子受体在免疫反应中发挥关键作用。在本研究中,在大口黑鲈(Micropterus salmoides)中鉴定出了五个JAKs家族成员(JAK1、JAK2a、JAK2b、JAK3和TYK2)。大口黑鲈的这五个JAKs成员包含几个保守的蛋白质结构域,包括一个FERM结构域、一个SH2结构域、一个假激酶(STyrkc)结构域和一个酪氨酸激酶(Tyrkc)结构域。系统发育分析表明,JAK2/2a/2b和JAK3形成一个进化枝,而JAK1/TYK2形成另一个单独的进化枝。实时定量PCR检测显示,所有五个JAKs基因在十个选定组织中组成性表达,在脾脏和头肾(HK)中高表达。在感染杀鱼爱德华氏菌后,这五个JAKs基因在感染后不同时间在脾脏和原代肝细胞中显著上调。进一步的选择压力分析表明,这五个JAKs成员在进化过程中经历了负选择压力。这五个JAK成员的FERM和Tyrkc结构域发生了纯化选择,并参与了与STAT3的相互作用,这通过蛋白质-蛋白质相互作用(PPI)网络和分子对接分析得到证实。此外,分子对接结果表明,氢键和盐键在JAK/STAT3复合物形成中起关键作用。我们的结果表明,JAKs在大口黑鲈对细菌的免疫反应中具有重要作用,为阐明鱼类中JAK/STAT信号通路的机制提供了依据。

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