Laboratory of Fish Biogenetics & Immune Evolution, College of Marine Science, Zhejiang Ocean University, Zhoushan, 316022, China; College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, 201306, China.
Laboratory of Fish Biogenetics & Immune Evolution, College of Marine Science, Zhejiang Ocean University, Zhoushan, 316022, China.
Fish Shellfish Immunol. 2018 Jun;77:1-7. doi: 10.1016/j.fsi.2018.02.003. Epub 2018 Feb 3.
MicroRNAs (miRNAs) are endogenous small non-coding RNAs that participate in the regulation of various biological processes. A series of microRNAs have been shown to be important regulators of both innate and adaptive immune responses, including RIG-I signaling pathway. In this study, we evaluated the regulation role of miR-210 in the RLRs signaling pathway of miiuy croaker. Upon poly(I:C) stimulation, the expression of miR-210 in both miiuy croaker spleen tissues and macrophages were significantly upregulated. By means of the dual luciferase reporter assay, a direct interaction between miR-210 and the 3-untranslated region (UTR) of Deubiquitinating enzyme A (DUBA) was confirmed, and we found that miR-210 could reduce the luciferase levels of wild-type DUBA 3'UTR, whereas mutant-type led to a complete abrogation of the negative effect. Furthermore, the negative regulatory effects of pre-miR-210 on DUBA have been indicated in a dose- and time-dependent manners. As DUBA is an important regulator involved in the RLRs signaling pathway and could bind with and regulate TRAF3, we also examined the expression patterns of DUBA and TRAF3 in vivo and in vitro. We found that the expression of both DUBA and TRAF3 were significantly changed upon poly(I:C) stimulation in miiuy croaker. The expression patterns between miR-210 and DUBA showed a negative correlation, which indicated that miR-210 can target and downregulate the expression of DUBA. Overall, these results will enrich the knowledge of immune response related miRNAs in miiuy croaker, which will be useful for better understanding the complicated regulatory networks in fish species.
微小 RNA（miRNAs）是参与调节各种生物过程的内源性小非编码 RNA。一系列 microRNAs 已被证明是先天和适应性免疫反应的重要调节剂，包括 RIG-I 信号通路。在这项研究中，我们评估了 miR-210 在米氏鲈 RLRs 信号通路中的调节作用。在 poly(I:C)刺激后，miR-210 在米氏鲈脾脏组织和巨噬细胞中的表达均显著上调。通过双荧光素酶报告基因检测，证实了 miR-210 与去泛素化酶 A（DUBA）的 3'非翻译区（UTR）之间的直接相互作用，并且我们发现 miR-210 可以降低野生型 DUBA 3'UTR 的荧光素酶水平，而突变型则完全消除了负效应。此外，miR-210 对 DUBA 的负调控作用呈剂量和时间依赖性。由于 DUBA 是 RLRs 信号通路中的重要调节剂，并且可以与 TRAF3 结合并调节 TRAF3，我们还在体内和体外检查了 DUBA 和 TRAF3 的表达模式。我们发现，在米氏鲈中，poly(I:C)刺激后 DUBA 和 TRAF3 的表达均发生显著变化。miR-210 和 DUBA 之间的表达模式呈负相关，表明 miR-210 可以靶向并下调 DUBA 的表达。总体而言，这些结果将丰富米氏鲈免疫反应相关 miRNAs 的知识，有助于更好地理解鱼类物种中复杂的调控网络。
