Luo Wen, Wu Hongling, Huang Ying, Zou Wenzheng, Han Fang
State Key Laboratory of Mariculture Breeding, Fisheries College, Jimei University, Xiamen 361000, PR China.
State Key Laboratory of Mariculture Breeding, Fisheries College, Jimei University, Xiamen 361000, PR China.
Comp Biochem Physiol B Biochem Mol Biol. 2025 Aug-Sep;279:111115. doi: 10.1016/j.cbpb.2025.111115. Epub 2025 Jun 5.
Viperin (Virus inhibitory protein, endoplasmic reticulum-associated, IFN-inducible) is a broad-spectrum antiviral protein widely involved in vertebrate innate immune regulation. This study cloned the 5' flanking promoter region (2010 bp) of the viperin gene (named as LcViperin) in large yellow croaker (Larimichthys crocea) and analyzed its promoter characteristics through bioinformatics and dual-luciferase reporter assays. Results indicated that the LcViperin promoter region contains two TATA boxes and several critical transcription factor binding sites, including interferon-stimulated response elements (ISREs), GATA-binding factor 1 (GATA1), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), interferon regulatory factor 7 (IRF7), and signal transduction and activator of transcription 1 (STAT1). Promoter region truncation experiments in HEK 293 T cells further confirmed that the core promoter region is located between -331 and - 121 bp. LcViperin overexpression significantly activated IRF3, IRF7, and IFN1 promoters in a dose-dependent manner. Domain deletion experiments demonstrated that the N-terminal and SAM (S-adenosyl-L-methionine) domains play key roles in promoter activation. Additionally, co-transfection with LcViperin and IRAK1 (interleukin-1 receptor-associated kinase 1, named as LcIRAK1) significantly enhanced IRF3, IRF7, and IFN1 promoter activity. This study reveals the regulatory characteristics of the large yellow croaker viperin promoter and its role in the interferon signaling pathway, providing a theoretical basis for understanding its immune regulatory mechanism and improving disease resistance in large yellow croaker. It also offers a scientific basis for understanding viperin regulation and its application in disease prevention in aquaculture.
蝰蛇毒素（病毒抑制蛋白，内质网相关，干扰素诱导）是一种广泛参与脊椎动物先天免疫调节的广谱抗病毒蛋白。本研究克隆了大黄鱼（Larimichthys crocea）中蝰蛇毒素基因（命名为LcViperin）的5'侧翼启动子区域（2010 bp），并通过生物信息学和双荧光素酶报告基因检测分析了其启动子特征。结果表明，LcViperin启动子区域包含两个TATA盒和几个关键转录因子结合位点，包括干扰素刺激反应元件（ISREs）、GATA结合因子1（GATA1）、活化B细胞核因子κ轻链增强子（NF-κB）、干扰素调节因子7（IRF7）和信号转导及转录激活因子1（STAT1）。在HEK 293 T细胞中进行的启动子区域截短实验进一步证实，核心启动子区域位于-331至-121 bp之间。LcViperin的过表达以剂量依赖的方式显著激活了IRF3、IRF7和IFN1启动子。结构域缺失实验表明，N端和SAM（S-腺苷-L-甲硫氨酸）结构域在启动子激活中起关键作用。此外，LcViperin与IRAK1（白细胞介素-1受体相关激酶1，命名为LcIRAK1）共转染显著增强了IRF3、IRF7和IFN1启动子活性。本研究揭示了大黄鱼蝰蛇毒素启动子的调控特征及其在干扰素信号通路中的作用，为理解其免疫调节机制和提高大黄鱼抗病能力提供了理论基础。它也为理解蝰蛇毒素调控及其在水产养殖疾病预防中的应用提供了科学依据。
