College of Animal Sciences, Zhejiang University, Hangzhou, China.
PLoS Pathog. 2022 Nov 16;18(11):e1010938. doi: 10.1371/journal.ppat.1010938. eCollection 2022 Nov.
Baculoviruses are virulent pathogens that infect a wide range of insects. They initiate infections via specific interactions between the structural proteins on the envelopes of occlusion-derived virions (ODVs) and the midgut cell surface receptors in hosts. However, host factors that are hijacked by baculoviruses for efficient infection remain largely unknown. In this study, we identified a membrane-associated protein sucrose hydrolase (BmSUH) as an ODV binding factor during Bombyx mori nucleopolyhedrovirus (BmNPV) primary infection. BmSUH was specifically expressed in the midgut microvilli where the ODV-midgut fusion happened. Knockout of BmSUH by CRISPR/Cas9 resulted in a significantly higher survival rate after BmNPV orally infection. Liquid chromatography-tandem mass spectrometry analysis and co-immunoprecipitation analysis demonstrated that PIF protein complex required for ODV binding could interact with BmSUH. Furthermore, fluorescence dequenching assay showed that the amount of ODV binding and fusion to the midgut decreased in BmSUH mutants compared to wild-type silkworm, suggesting the role of BmSUH as an ODV binding factor that mediates the ODV entry process. Based on a multilevel survey, the data showed that BmSUH acted as a host factor that facilitates BmNPV oral infection. More generally, this study indicated that disrupting essential protein-protein interactions required for baculovirus efficient entry may be broadly applicable to against viral infection.
杆状病毒是一种烈性病原体,可感染多种昆虫。它们通过包膜出芽型病毒粒子(ODV)表面的结构蛋白与宿主中肠细胞表面受体之间的特异性相互作用引发感染。然而,杆状病毒劫持宿主因子以实现有效感染的机制在很大程度上仍不清楚。在本研究中,我们鉴定了一种膜相关蛋白蔗糖水解酶(BmSUH),它是家蚕核型多角体病毒(BmNPV)初次感染过程中 ODV 的结合因子。BmSUH 在 ODV 与中肠融合发生的中肠微绒毛中特异性表达。通过 CRISPR/Cas9 敲除 BmSUH 会导致 BmNPV 经口感染后的存活率显著提高。液相色谱-串联质谱分析和共免疫沉淀分析表明,ODV 结合所需的 PIF 蛋白复合物可以与 BmSUH 相互作用。此外,荧光淬灭测定表明,与野生型家蚕相比,ODV 结合和与中肠融合的量在 BmSUH 突变体中减少,表明 BmSUH 作为一种 ODV 结合因子,介导 ODV 进入过程。基于多层次调查,数据表明 BmSUH 作为一种宿主因子,促进了 BmNPV 的经口感染。更普遍地说,这项研究表明,破坏杆状病毒有效进入所需的关键蛋白-蛋白相互作用可能广泛适用于对抗病毒感染。