Molecular cloning, expression and functional analysis of Atg16L1 from orange-spotted grouper (Epinephelus coioides)
Molecular cloning, expression and functional analysis of Atg16L1 from orange-spotted grouper (Epinephelus coioides)
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斜带石斑鱼Atg16L1的分子克隆、表达及功能分析
DOI：
10.1016/j.fsi.2019.09.004
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发表时间：
2019-11-01
影响因子：
4.7
通讯作者：
Qin, Qiwei
中科院分区：
文献类型：
--
作者：
Li, Chen;Wang, Liqun;Qin, Qiwei
自噬相关基因16（Atg16）是自噬体形成的核心蛋白，参与哺乳动物的自噬活动、泛素蛋白酶体系统和炎症反应。在这项研究中，我们克隆和分析了一个Atg16同源物从橙点石斑鱼（Epineoparus coioides）（EcAtg16L1）。EcAtg16L1编码656个氨基酸，与大黄鱼（Larimichthys crocea）和人（Homo sapiens）的同源性分别为94.22%和72.65%。EcAtg16L1含有一个保守的Atg16结构域和一个WD重复序列结构域。亚细胞定位结果表明，EcAtg16L1在石斑鱼细胞质中呈点状分布。EcAtg 16 L1过表达促进新加坡石斑鱼虹彩病毒（SGIV）和红斑石斑鱼神经坏死病毒（RGNNV）的复制，这通过病毒基因转录和病毒外壳蛋白的增加来证明。此外，EcAtg16L1过表达负调节干扰素（IFN）相关分子和促炎细胞因子，并降低IFN，IFN刺激的反应元件，和核因子xB启动子的活动。综上所述，除了其在自噬体形成中的功能之外，EcAtg16L1还在促进SGIV和RGNNV复制中起作用，并且前病毒作用可能涉及其对干扰素和炎症反应的下调。
Autophagy related gene 16 (Atg16), which encodes a core protein for autophagosome formation, participates in autophagy activity, the ubiquitin proteasome system and inflammatory response in mammals. In this study, we cloned and characterized an Atg16 homolog from orange-spotted grouper (Epinephelus coioides) (EcAtg16L1). EcAtg16L1 encodes a 656-amino acid polypeptide, which shares 94.22% and 72.65% homology with large yellow croakers (Larimichthys crocea) and humans (Homo sapiens), respectively. EcAtg16L1 contains a conserved Atg16 domain and a WD-repeat-containing domain. Subcellular localization showed that EcAtg16L1 was distributed in the cytoplasm of grouper cells with a dot-like pattern. EcAtg16L1 overexpression promoted Singapore grouper iridovirus (SGIV) and red-spotted grouper nervous necrosis virus (RGNNV) replication, as evidenced by the increase in viral gene transcription and viral coat protein. Furthermore, EcAtg16L1 overexpression negatively regulated interferon (IFN)-related molecules and proinflammatory cytokines, and decreased IFN, IFN-stimulated response element, and nuclear factor xB promoter activities. Taken together, aside from its function in autophagosome formation, EcAtg16L1 also plays role in promoting SGIV and RGNNV replication and the proviral effect might involve its down regulation to interferon and inflammatory responses.