College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, Xinjiang, China.
College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, Xinjiang, China.
Vet Microbiol. 2018 Feb;215:43-48. doi: 10.1016/j.vetmic.2018.01.002. Epub 2018 Jan 9.
MicroRNAs (miRNAs) are endogenous ∼22 nt noncoding RNAs that control the translation initiation and stability of target genes in a sequence-specific manner and, thus, play important regulatory roles in animals and plants. Homologs of Dom34, called Pelota or PELO, are broadly conserved in eukaryotes and archaea. Biochemical and genetic studies indicate that eukaryotic Dom34/Pelota plays an important role in cell division, differentiation of germline stem cells, and stem cell self-renewal by controlling the expression of specific genes at the translational level. Additionally, it is reported that Pelota is specifically required for high efficiency synthesis of proteins in numerous viruses. In earlier studies, we found the Bos taurus bta-miR-2411 (shortly miR-2411 herein) was significantly upregulated by more than 2.1 times in bovine viral diarrhea virus (BVDV) strain NADL-infected Madin-Darby bovine kidney (MDBK) cells after 8 h post-infection (pi) compared to normal MDBK cells without BVDV infection. Moreover, miR-2411 overexpression significantly reduced the BVDV E1 mRNA level and viral titer. Nevertheless, the mechanisms of miR-2411 attenuating on viral replication remain unclear. Here, we report that miR-2411 as a novel microRNA regulates BVDV NADL replication via directly targeting the Pelota gene in MDBK cells. We investigated whether the potential target sequences of miR-2411, located in the Pelota 3'UTR, and miR-2411 agomir transfection attenuated Pelota mRNA and protein levels. Indeed, upon miR-2411 overexpression, BVDV NADL replication was prevented. Importantly, BVDV NADL replication levels were reversed to normal levels as a result of the Pelota rescuing experiment even though miR-2411 was existent. Overall, we profiled the unique role of miR-2411 in regulating BVDV NADL replication and provided a novel strategy for generalized inhibition of viral infection.
微小 RNA（miRNAs）是内源性的约 22 个核苷酸的非编码 RNA，以序列特异性方式控制靶基因的翻译起始和稳定性，因此在动物和植物中发挥重要的调节作用。与 Dom34 同源的蛋白，称为 Pelota 或 PELO，在真核生物和古菌中广泛保守。生化和遗传研究表明，真核生物 Dom34/Pelota 通过在翻译水平上控制特定基因的表达，在细胞分裂、生殖干细胞分化和干细胞自我更新中发挥重要作用。此外，有报道称 Pelota 是许多病毒高效合成蛋白质所必需的。在早期的研究中，我们发现牛病毒性腹泻病毒（BVDV）株 NADL 感染后的牛肾细胞（MDBK）中，牛 taur miR-2411（简称 miR-2411）在感染后 8 小时的表达水平比未感染 BVDV 的正常 MDBK 细胞显著上调了 2.1 倍以上。此外，miR-2411 的过表达显著降低了 BVDV E1 mRNA 水平和病毒滴度。然而，miR-2411 抑制病毒复制的机制尚不清楚。在这里，我们报告 miR-2411 作为一种新型 microRNA，通过直接靶向 MDBK 细胞中的 Pelota 基因来调节 BVDV NADL 的复制。我们研究了 miR-2411 的潜在靶序列，位于 Pelota 3'UTR 中，以及 miR-2411 agomir 转染是否能减弱 Pelota mRNA 和蛋白水平。事实上，当 miR-2411 过表达时，BVDV NADL 的复制被阻止。重要的是，尽管存在 miR-2411，Pelota 的挽救实验使 BVDV NADL 的复制水平恢复到正常水平。总的来说，我们描绘了 miR-2411 在调节 BVDV NADL 复制中的独特作用，并为普遍抑制病毒感染提供了一种新策略。
