Inhibition of miR-274-3p increases BmCPV replication by regulating the expression of BmCPV NS5 gene in Bombyx mori

Inhibition of miR-274-3p increases BmCPV replication by regulating the expression of BmCPV NS5 gene in Bombyx mori
复制标题

抑制 miR-274-3p 通过调节家蚕 BmCPV NS5 基因的表达来增加 BmCPV 复制

DOI:
10.1007/s11262-017-1466-7
复制
发表时间:
2017-05
期刊:
影响因子:
1.6
通讯作者:
Xijie Guo
Xijie Guo
中科院分区:
医学4区
文献类型:
--
作者:
Ping Wu;Xiaoxu Jiang;Qi Shang;Annan Enoch;Tao Cheng;Xijie Guo

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家蚕细胞质多角体病毒(BmCPV)是给蚕桑业带来巨大挑战的主要病原之一。越来越多的证据表明,microRNA 在昆虫宿主与病原体相互作用的调节中发挥着关键作用。在家蚕中发现了 MicroRNA,但它们是否以及如何影响家蚕与 BmCPV 的相互作用仍不清楚。在这里,我们研究了 miR-274-3p 对 BmCPV 感染的蚕幼虫中 BmCPV 复制的影响。在我们的研究中,基于生物信息学分析和荧光素酶报告基因测定,BmCPV 非结构蛋白 5 (NS5) 被确定为 miR-274-3p 的靶标。根据 qRT-PCR 和蛋白质印迹结果,在 miR-274-3p 抑制剂存在下,NS5 的丰度显着增加。此外,qRT-PCR结果显示,与阴性对照幼虫相比,施用miR-274-3p抑制剂后,幼虫中BmCPV多角体蛋白基因的表达显着增加。我们的结果表明,抑制 miR-274-3p 可以通过上调 BmCPV NS5 基因表达来促进 BmCPV 复制,对于进一步探索家蚕和 BmCPV 之间的相互作用具有重要意义。
Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) is one of the major pathogens that pose a big challenge to the sericulture industry. Growing evidences have shown that microRNAs play key roles in the regulations of host-pathogen interactions in insects. MicroRNAs have been found in silkworms, whether and how they affect the silkworm-BmCPV interactions are still unknown. Here we investigate the effect of miR-274-3p on the BmCPV replication in the BmCPV-infected silkworm larvae. In our study, BmCPV Nonstructural protein 5 (NS5) was identified to be the target of miR-274-3p based on bioinformatics analysis and luciferase reporter assay. The abundance of NS5 was significantly increased in the presence of miR-274-3p inhibitor based on the qRT-PCR and Western blotting results. Further, qRT-PCR results revealed that the expression of polyhedrin gene of BmCPV in the larvae after applying miR-274-3p inhibitor was significantly increased comparing with that of larvae with negative control. Our results suggest that inhibition of miR-274-3p could facilitate BmCPV replication by up-regulating BmCPV NS5 gene expression and are insightful for further exploring the interactions between silkworm and BmCPV.
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