1State Key Laboratory of Agrobiotechnology and Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Biological Sciences, China Agricultural University, Beijing 100193, PR China.
2Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, PR China.
J Gen Virol. 2018 Dec;99(12):1671-1680. doi: 10.1099/jgv.0.001166. Epub 2018 Nov 1.
Porcine reproductive and respiratory syndrome (PRRS) is one of the most important diseases in pigs. MicroRNAs (miRNAs) have emerged as an important regulator of virus-host cell interactions and miR-30c has been found to facilitate PRRSV replication. Here, we found that the interferon-alpha/beta receptor beta chain (IFNAR2) was down-regulated, while miR-30c was up-regulated during HV (a highly pathogenic type 2 PRRSV strain) and CH-1a (a classic type 2 PRRSV strain) infection. Subsequently, using bioinformatics analysis, we predicted that the IFNAR2 was targeted by miR-30c. A luciferase assay verified that the 3' UTR of IFNAR2 was targeted by miR-30c, as a mutation on either the target sequence or the miR-30c seed sequence reversed the luciferase activity. In addition, miR-30c and IFNAR2 mRNA were physically co-localized in RNA-induced silencing complex (RISC). Importantly, we showed that miR-30c also impaired the induction of IFN-stimulated genes (ISGs) by targeting IFNAR2. Our findings further reveal the mechanism of miR-30c promoting PRRSV replication.
猪繁殖与呼吸综合征（PRRS）是猪的最重要疾病之一。微小 RNA（miRNA）已成为病毒-宿主细胞相互作用的重要调节剂，并且已经发现 miR-30c 有助于 PRRSV 复制。在这里，我们发现干扰素-α/β受体β链（IFNAR2）在 HV（一种高致病性 2 型 PRRSV 株）和 CH-1a（一种经典 2 型 PRRSV 株）感染期间下调，而 miR-30c 则上调。随后，通过生物信息学分析，我们预测 IFNAR2 是 miR-30c 的靶标。荧光素酶报告基因实验验证了 IFNAR2 的 3'UTR 是由 miR-30c 靶向的，因为靶序列或 miR-30c 种子序列上的突变逆转了荧光素酶活性。此外，miR-30c 和 IFNAR2 mRNA 在 RNA 诱导沉默复合物（RISC）中物理共定位。重要的是，我们表明 miR-30c 通过靶向 IFNAR2 也损害了 IFN 刺激基因（ISGs）的诱导。我们的研究结果进一步揭示了 miR-30c 促进 PRRSV 复制的机制。
