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家蚕肌动蛋白相互作用蛋白胸腺素的过表达和抑制表达对家蚕核型多角体病毒增殖和复制的影响。

Effects of overexpression and inhibited expression of thymosin, an actin-interacting protein from Bombyx mori, on BmNPV proliferation and replication.

作者信息

Liao Jinxu, Zhang Chen, Ru Wenjuan, Wang Dan, Zhang Wenping

机构信息

College of Life Science, Institute of Biochemistry, Zhejiang Sci-Tech University, Hangzhou, Zhejiang, China.

Zhejiang Provincial Key Laboratory of Silkworm Bioreactor and Biomedicine, Hangzhou, Zhejiang, China.

出版信息

Arch Insect Biochem Physiol. 2018 May;98(1):e21449. doi: 10.1002/arch.21449. Epub 2018 Jan 29.

Abstract

Previous study showed that exogenously applied recombinant thymosin from Bombyx mori (BmTHY) reduces B. mori nucleopolyhedrovirus (BmNPV) proliferation in silkworm. Which stands to reason that BmTHY in B. mori is crucial for the defense against BmNPV. However, little is known about the effect of endogenously overexpressed or repressed BmTHY on B. mori resistance to virus infection. To study this issue, we constructed an overexpression and inhibited expression systems of BmTHY in BmN cells. The viral titer and the analysis from the quantitative real-time polymerase chain reaction (PCR) revealed that overexpression of BmTHY decreased the copies of BmNPV gene gp41, which goes over to inhibit the proliferation of BmNPV in BmN cells, while the inhibited expression of BmTHY significantly enhanced viral proliferation in infected BmN cells. These results indicated that endogenous BmTHY can inhibit BmNPV proliferation and replication in infected BmN cells. Furthermore, Co-IP showed that BmTHY could bind to actin in BmN cells. Also, the overexpression or inhibited expression of BmTHY shifted the ratio of F/G-actin in infected BmN cells. Lastly, the BmTHY, an actin-interacting protein, might be one of the key host factors against BmNPV, which inhibits viral proliferation and replication in BmN cells.

摘要

先前的研究表明,外源应用的家蚕重组胸腺素(BmTHY)可减少家蚕核型多角体病毒(BmNPV)在家蚕中的增殖。由此可以推断,家蚕体内的BmTHY对于抵御BmNPV至关重要。然而,关于内源性过表达或抑制表达BmTHY对家蚕抗病毒感染能力的影响,人们知之甚少。为了研究这个问题,我们构建了BmTHY在家蚕BmN细胞中的过表达和抑制表达系统。病毒滴度以及定量实时聚合酶链反应(PCR)分析表明,BmTHY的过表达降低了BmNPV基因gp41的拷贝数,进而抑制了BmNPV在BmN细胞中的增殖,而BmTHY的抑制表达则显著增强了病毒在受感染BmN细胞中的增殖。这些结果表明,内源性BmTHY可以抑制BmNPV在受感染BmN细胞中的增殖和复制。此外,免疫共沉淀(Co-IP)显示BmTHY可以与BmN细胞中的肌动蛋白结合。而且,BmTHY的过表达或抑制表达改变了受感染BmN细胞中F型/ G型肌动蛋白的比例。最后,BmTHY作为一种与肌动蛋白相互作用的蛋白,可能是家蚕抵御BmNPV的关键宿主因子之一,它可以抑制病毒在BmN细胞中的增殖和复制。

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