Lentivirus-mediated Bos taurus bta-miR-29b overexpression interferes with bovine viral diarrhoea virus replication and viral infection-related autophagy by directly targeting ATG14 and ATG9A in Madin-Darby bovine kidney cells
Lentivirus-mediated Bos taurus bta-miR-29b overexpression interferes with bovine viral diarrhoea virus replication and viral infection-related autophagy by directly targeting ATG14 and ATG9A in Madin-Darby bovine kidney cells
复制标题
慢病毒介导的牛bta-miR-29b过表达通过直接靶向Madin-Darby牛肾细胞中的ATG14和ATG9A干扰牛病毒性腹泻病毒复制和病毒感染相关的自噬
DOI：
10.1099/vir.0.067140-0
复制
发表时间：
2015-01-01
影响因子：
3.8
通讯作者：
Chen, Chuangfu
中科院分区：
文献类型：
--
作者：
Fu, Qiang;Shi, Huijun;Chen, Chuangfu
MicroRNAs （miRNAs）是一类短的内源性RNA分子，通过与靶基因3′utr上部分互补的位点配对，能够控制真核生物的发育、自噬、细胞凋亡和应激反应。最近的研究表明，mirna在宿主病原体相互作用的复杂网络中起着关键的效应作用。值得注意的是，我们发现在感染牛病毒性腹泻病毒（BVDV）菌株nadl感染的Madin Darby牛肾（MDBK）细胞6小时后，与正常的MDBK细胞相比，Boa taurus bta-miR-29b（本文简称miR-29b）显著上调>2.3倍。然而，miR-29b在BVDV感染及其发病机制中的作用尚不清楚。在这里，我们报道了miR-29b对BVDV NADL复制和病毒感染相关自噬的抑制作用。miRNA前体表达慢病毒介导miR-29b过表达，通过直接下调两种关键自噬相关蛋白ATG14和ATG9A的细胞内表达水平，导致BVDV NADL感染相关自噬的衰减。此外，ATG14和ATG9A过表达挽救不仅逆转了mir -29b抑制的自噬，而且增加了BVDV NADL的复制。在之前的研究中，我们发现MDBK细胞早期的自噬促进了BVDV NADL的复制，抑制自噬抑制了BVDV NADL的复制，本研究也证实了这一点。总之，我们的研究结果建立了miR-29b与病毒复制之间的新联系，也为宿主细胞与病原体之间的密切相互作用提供了新的途径。
MicroRNAs (miRNAs) are a class of short endogenous RNA molecules with the ability to control development, autophagy, apoptosis and the stress response in eukaryotes by pairing with partially complementary sites in the 3' UTRs of targeted genes. Recent studies have demonstrated that miRNAs serve as critical effectors in intricate networks of host pathogen interactions. Notably, we found that Boa taurus bta-miR-29b (referred to as miR-29b herein) was significantly upregulated >2.3-fold in bovine viral diarrhoea virus (BVDV) strain NADL-infected Madin Darby bovine kidney (MDBK) cells 6 h post-infection compared with normal MDBK cells. However, the roles of miR-29b in BVDV infection and pathogenesis remain unclear. Here, we report the inhibitory effects of miR-29b on BVDV NADL replication and viral infection-related autophagy. miR-29b overexpression mediated by miRNA precursor-expressing lentivirus resulted in the attenuation of BVDV NADL infection-related autophagy by directly downregulating the intracellular expression levels of two key autophagy-associated proteins, ATG14 and ATG9A. Moreover, ATG14 and ATG9A overexpression rescue not only reversed miR-29b-inhibited autophagy, but also increased BVDV NADL replication. In previous studies, we found that the early stages of autophagy contributed to BVDV NADL replication in MDBK cells and that the inhibition of autophagy repressed BVDV NADL replication, which was also proved in the present study. Collectively, our results establish a novel link between miR-29b and viral replication, and also provide a new pathway for the intimate interaction between host cells and pathogens.