Molecular cloning and characterization of FADD from the orange-spotted grouper ( Epinephelus coioides )
Molecular cloning and characterization of FADD from the orange-spotted grouper ( Epinephelus coioides )
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橙斑石斑鱼 (Epinephelus coioides) FADD 的分子克隆和表征
DOI：
10.1016/j.fsi.2018.01.018
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发表时间：
2018
影响因子：
4.7
通讯作者：
Qiwei Qin
中科院分区：
文献类型：
--
作者：
Xin Zhang;Shaoqing Zang;Chen Li;Jingguang Wei;Qiwei Qin
具有死亡结构域的 Fas 相关蛋白 (FADD) 是传递由主要死亡受体介导的细胞凋亡信号的关键接头蛋白。除了作为细胞死亡的重要工具外，FADD 还与增殖、细胞周期进展、肿瘤发展、炎症、先天免疫和自噬有关。在本研究中，克隆了来自橙点石斑鱼（Epinephelus coioides）的 FADD 同源物（EcFADD），并分析了其在鱼类免疫中的可能作用。 EcFADD的全长cDNA包含808个碱基对(bp)，其中包括一个573bp的开放阅读框，编码190个氨基酸的蛋白质，预测分子量为21.81kDa。实时定量聚合酶链反应分析表明 EcFADD 分布在所有检查的组织中。 EcFADD在E的脾脏中的表达。当受到新加坡石斑鱼虹彩病毒 (SGIV) 或聚肌苷-聚胞苷酸 (poly[I:C]) 攻击时，coioides 的表达存在差异性上调。 EcFADD 大量分布在石斑鱼脾 (GS) 和黑头鲦鱼 (FHM) 上皮细胞的细胞质和细胞核中。 EcFADD 的过度表达抑制 SGIV 感染和复制以及 SGIV 诱导的细胞凋亡。为了实现抗病毒和抗凋亡活性，FADD促进抗病毒IFN信号通路中干扰素刺激反应元件（ISRE）和I型干扰素（IFN）基因的激活，并抑制凋亡相关转录因子p53的激活。我们的结果不仅表征了 FADD，还揭示了新的免疫功能以及 FADD 响应病毒感染和病毒诱导的细胞凋亡的分子机制。
Fas-associated protein with death domain (FADD) is the key adaptor protein that transmits apoptotic signals mediated by the main death receptors. Besides being an essential instrument in cell death, FADD is also implicated in proliferation, cell cycle progression, tumor development, inflammation, innate immunity, and autophagy. In the present study, a FADD homologue (EcFADD) from the orange-spotted grouper (Epinephelus coioides) was cloned and its possible role in fish immunity was analyzed. The full length cDNA of EcFADD contains 808 base pairs (bp), including a 573 bp open reading frame that encodes a 190 amino acid protein with a predicted molecular mass of 21.81 kDa. Quantitative real-time polymerase chain reaction analysis indicated that EcFADD was distributed in all examined tissues. The expression of EcFADD in the spleen ofE. coioideswas differentially up-regulated when challenged with Singapore grouper iridovirus (SGIV) or polyinosine-polycytidylic acid(poly[I:C]). EcFADD was abundantly distributed in both the cytoplasm and nucleus in grouper spleen (GS) and fathead minnow (FHM) epithelial cells. Over-expression of EcFADD inhibited SGIV infection and replication and SGIV-induced apoptosis. To achieve antiviral and anti-apoptosis activities, FADD promoted the activation of interferon-stimulated response element (ISRE) and type I interferon (IFN) genes in the antiviral IFN signaling pathway and inhibited activation of apoptosis-related transcription factors p53. Our results not only characterize FADD but also reveal new immune functions and the molecular mechanisms by which FADD responds to virus infection and virus-induced apoptosis.