College of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication), State Key Laboratory of Marine Resource Utilization in South China Sea, Hainan Aquaculture Breeding Engineering Research Center, Hainan Academician Team Innovation Center, Hainan University, Haikou, 570228, China.
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, The Innovation Academy of Seed Design, Chinese Academy of Sciences, Wuhan, 430072, China.
Fish Shellfish Immunol. 2023 Dec;143:109163. doi: 10.1016/j.fsi.2023.109163. Epub 2023 Oct 13.
The golden pompano (Trachinotus blochii), a pivotal commercial marine species in China, has gained significant popularity worldwide. However, accompanied with rapid growth and high density aquaculture, golden pompano has been seriously threatened by Nervous necrosis virus (NNV), while its molecular biology research regarding the innate immune system remains unexplored, which is crucial for understanding the activation of interferon (IFN) production and antiviral responses. In this study, we aimed to identify the characterization and function of golden pompano TANK-binding kinase 1 (gpTBK1), thereby providing evidence of the conservation of this classical factor in the RLR pathway among marine fish. Initially, we found the expression of gpTBK1 upregulation in diseased golden pompano with NNV infection and we successfully cloned the full-length open reading frame (ORF) of gpTBK1, consisting of 2172 nucleotides encoding 723 amino acids, from the head kidney. Subsequent analysis of the amino acid sequence revealed homology between gpTBK1 and other fish TBK1 proteins, with conserved N-terminal Serine/Threonine protein kinases catalytic domain (S_TKc) and C-terminal coiled coil domain (CCD). Moreover, the expression pattern showed that gpTBK1 exhibited ubiquitous expression across all evaluated tissues. Furthermore, functional identification experiments indicated that gpTBK1 activated interferon promoters' activity in golden pompano and induced the expression of downstream IFN-stimulated genes (ISGs). Notably, gpTBK1 was found to co-localize and interact with gpIRF3 in the cytoplasm. Collectively, these data provide a comprehensive analysis of the characterization and functional role of gpTBK1 in promoting interferon production. This research may facilitate the further study of the innate antiviral response, particularly the anti-NNV mechanisms, in golden pompano.
金黄鰤（Trachinotus blochii），中国重要的商业海水养殖鱼类之一，在全球范围内广受欢迎。然而，随着其养殖业的快速发展和高密度养殖，金黄鰤受到神经坏死病毒（Nervous necrosis virus，NNV）的严重威胁，但其先天免疫系统的分子生物学研究仍未被探索，这对于了解干扰素（IFN）产生和抗病毒反应的激活至关重要。在本研究中，我们旨在鉴定金黄鰤 TANK 结合激酶 1（gpTBK1）的特征和功能，从而为海洋鱼类 RLR 途径中这一经典因子的保守性提供证据。最初，我们发现患有 NNV 感染的患病金黄鰤中 gpTBK1 的表达上调，并且我们成功地从头肾克隆了 gpTBK1 的全长开放阅读框（ORF），由编码 723 个氨基酸的 2172 个核苷酸组成。随后对氨基酸序列的分析表明，gpTBK1 与其他鱼类 TBK1 蛋白具有同源性，具有保守的 N 端丝氨酸/苏氨酸蛋白激酶催化结构域（S_TKc）和 C 端卷曲螺旋结构域（CCD）。此外，表达模式表明 gpTBK1 在所有评估的组织中均具有广泛表达。此外，功能鉴定实验表明，gpTBK1 激活了金黄鰤干扰素启动子的活性并诱导了下游干扰素刺激基因（ISGs）的表达。值得注意的是，gpTBK1 被发现与细胞质中的 gpIRF3 共定位和相互作用。总之，这些数据提供了 gpTBK1 促进干扰素产生的特征和功能作用的全面分析。这项研究可能有助于进一步研究金黄鰤的先天抗病毒反应，特别是抗 NNV 机制。
