Identification and Functional Analysis of BmNPV-Interacting Proteins From Bombyx mori (Lepidoptera) Larval Midgut Based on Subcellular Protein Levels
Identification and Functional Analysis of BmNPV-Interacting Proteins From Bombyx mori (Lepidoptera) Larval Midgut Based on Subcellular Protein Levels
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DOI：
10.3389/fmicb.2020.01481
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发表时间：
2020-06
影响因子：
5.2
通讯作者：
Shang-zhi Zhang;Lin-bao Zhu;Dong Yu;Ling-ling You;Jie Wang;Hui-hua Cao;Ying-xue Liu;Yu-ling Wang-Yu
中科院分区：
文献类型：
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作者：
Shang-zhi Zhang;Lin-bao Zhu;Dong Yu;Ling-ling You;Jie Wang;Hui-hua Cao;Ying-xue Liu;Yu-ling Wang-Yu
家蚕核型多角体病毒（BmNPV）是造成严重经济损失的主要病原。然而，家蚕抗BmNPV的分子机制以及该病毒在感染过程中与宿主的相互作用仍不清楚。为探索家蚕病毒结合蛋白，采用单向、双向电泳和远蛋白质印迹结合质谱技术，对家蚕中肠亚细胞组分中可能与BmNPV相互作用的蛋白质进行了体外分析。共24种蛋白质被确定为在两个亚细胞级分（线粒体和微粒体）中特异性结合到出芽病毒（BV）。这些蛋白参与病毒的运输、能量代谢、细胞凋亡和病毒增殖等过程，在不同抗性株系中的表达谱不同。特别是，几乎所有鉴定的蛋白质在BmNPV感染后的A35株中下调。有趣的是，在家蚕中肠的胞质组分中没有发现病毒结合蛋白。两个候选蛋白，RACK 1和VDAC 2，与BV的相互作用，确定与远蛋白质印迹和反向远蛋白质印迹。我们推测与病毒相互作用的蛋白质可以增强或抑制病毒的感染。这些数据为进一步研究寄主与BmNPV的相互作用提供了全面有用的信息。
Bombyx mori nucleopolyhedrovirus (BmNPV) is a major pathogen causing severe economic loss. However, the molecular mechanism of silkworm resistance to BmNPV and the interactions of this virus with the host during infection remain largely unclear. To explore the virus-binding proteins of silkworms, the midgut subcellular component proteins that may interact with BmNPV were analyzed in vitro based on one- and two-dimensional electrophoresis and far-western blotting combined with mass spectrometry (MS). A total of 24 proteins were determined to be specifically bound to budded viruses (BVs) in two subcellular fractions (mitochondria and microsomes). These proteins were involved in viral transportation, energy metabolism, apoptosis and viral propagation, and they responded to BmNPV infection with different expression profiles in different resistant strains. In particular, almost all the identified proteins were downregulated in the A35 strain following BmNPV infection. Interestingly, there were no virus-binding proteins identified in the cytosolic fraction of the silkworm midgut. Two candidate proteins, RACK1 and VDAC2, interacted with BVs, as determined with far-western blotting and reverse far-western blotting. We speculated that the proteins interacting with the virus could either enhance or inhibit the infection of the virus. The data provide comprehensive useful information for further research on the interaction of the host with BmNPV.