miR-142a-3p promotes the proliferation of porcine hemagglutinating encephalomyelitis virus by targeting Rab3a
miR-142a-3p promotes the proliferation of porcine hemagglutinating encephalomyelitis virus by targeting Rab3a
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miR-142a-3p通过靶向Rab3a促进猪血凝性脑脊髓炎病毒增殖
DOI：
10.1007/s00705-019-04470-z
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发表时间：
2019
影响因子：
2.7
通讯作者：
Zi Li
中科院分区：
文献类型：
--
作者：
Peng Fan;Jiyu Guan;Wenqi He;Xiaoling Lv;Shiyu Hu;Yungang Lan;Kui Zhao;Feng Gao;Fang Li;Gencheng Fan;Hongbin He;Zi Li
猪血凝性脑脊髓炎病毒（PHEV）是一种典型的嗜神经冠状病毒，主要侵入仔猪中枢神经系统，引起仔猪呕吐和消瘦。新出现的证据表明，PHEV改变了microRNA（miRNA）的表达谱，miRNA也被假定参与其发病机制，但这一过程的机制尚未得到充分探讨。在这项研究中，我们发现PHEV感染上调了N2 a细胞和小鼠CNS中miR-142 a-3 p RNA的表达。通过miRNA抑制剂下调miR-142 a-3 p导致病毒增殖的显著抑制，这意味着它充当PHEV增殖的正调节剂。使用双荧光素酶报告基因分析，发现miR-142 a-3 p直接结合Rab 3a mRNA的3'非翻译区（3' UTR）并下调其表达。通过用miR-142 a-3 p模拟物或Rab 3a siRNA转染来敲低Rab 3a表达显著增加了N2 a细胞中的PHEV复制。相反，miR-142 a-3 p抑制剂的使用或Rab 3a的过表达导致在mRNA和蛋白质水平上对病毒产生的显著限制。我们的数据表明，miR-142 a-3 p通过直接靶向Rab 3a mRNA促进PHEV增殖，这为PHEV相关发病机制和病毒-宿主相互作用的机制提供了新的见解。
Porcine hemagglutinating encephalomyelitis virus (PHEV) is a typical neurotropic coronavirus that mainly invades the central nervous system (CNS) in piglets and causes vomiting and wasting disease. Emerging evidence suggests that PHEV alters microRNA (miRNA) expression profiles, and miRNA has also been postulated to be involved in its pathogenesis, but the mechanisms underlying this process have not been fully explored. In this study, we found that PHEV infection upregulates miR-142a-3p RNA expression in N2a cells and in the CNS of mice. Downregulation of miR-142a-3p by an miRNA inhibitor led to a significant repression of viral proliferation, implying that it acts as a positive regulator of PHEV proliferation. Using a dual-luciferase reporter assay, miR-142a-3p was found to bind directly bound to the 3’ untranslated region (3’UTR) of Rab3a mRNA and downregulate its expression. Knockdown of Rab3a expression by transfection with an miR-142a-3p mimic or Rab3a siRNA significantly increased PHEV replication in N2a cells. Conversely, the use of an miR-142a-3p inhibitor or overexpression of Rab3a resulted in a marked restriction of viral production at both the mRNA and protein level. Our data demonstrate that miR-142a-3p promotes PHEV proliferation by directly targeting Rab3a mRNA, and this provides new insights into the mechanisms of PHEV-related pathogenesis and virus-host interactions.