Abstract：
Ube3a is a member of the E3 ubiquitin ligase HECTc family, and its role has been established in neurodevelopmental disorders. However, studies on its role in Japanese flounder are scarce. Thus, in this study, the ube3a of Japanese flounder was cloned, and its role in conferring resistance against Chinook salmon bafnivirus (CSBV) was analyzed. Japanese flounder ube3a encoded a protein containing 834 amino acids. Interestingly, its homology with the Atlantic halibut was determined to be 94%. In addition, there were differential expressions of ube3a in different tissues of Japanese flounder, with the highest expression level observed in the fin, followed by the gills and skin (P ≤ 0.05). Subcellular localization analysis revealed that Ube3a is a cytoplasmic protein. We established an in vitro CSBV infection model using Japanese flounder gill cell line (FG). After ube3a overexpression, the viral load was significantly lower than that of the control group (P ≤ 0.05). Contrastingly, after incubation of FG cells with an E3 ubiquitin ligase inhibitor, the viral load was significantly higher than in the control group (P ≤ 0.01). Then, the expression levels of nf-κb, traf3, and tnf-α after incubation with an E3 ubiquitin ligase inhibitor were examined. The results demonstrated that ube3a may exerted a significant antiviral effect in Japanese flounder via the ubiquitination pathway.
摘要：
Ube3a是E3泛素连接酶HECTc家族的成员，它在神经发育障碍中的作用已经确立。然而,关于其在日本比目鱼中的作用的研究很少。因此，在这项研究中，日本比目鱼的ube3a被克隆，并分析了其在赋予抗Chinook鲑鱼bafnivirus（CSBV）中的作用。日本比目鱼ube3a编码一种含有834个氨基酸的蛋白质。有趣的是,与大西洋大比目鱼的同源性确定为94％。此外,ube3a在比目鱼的不同组织中存在差异表达，在鳍中观察到最高的表达水平，其次是ill和皮肤（P≤0.05）。亚细胞定位分析显示Ube3a是细胞质蛋白。我们使用日本比目鱼g细胞系（FG）建立了体外CSBV感染模型。ube3a过表达后，病毒载量显著低于对照组(P≤0.05)。相反，FG细胞与E3泛素连接酶抑制剂孵育后，病毒载量显著高于对照组(P≤0.01)。然后，NF-κB的表达水平，检查了与E3泛素连接酶抑制剂孵育后的traf3和tnf-α。结果表明，ube3a可能通过泛素化途径在日本比目鱼中发挥显着的抗病毒作用。
