Ding Xin-Yi, Su Jiang-Zhi, Yu Wen-Jing, Jiao Qian-Qian, Shao Zuo-Min, Wan Ai-Jun, Qin Sheng, Sun Xia, Wang Xue-Yang, Lü Ding-Ding, Li Mu-Wang
Jiangsu Key Laboratory of Sericultural and Animal Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, Jiangsu, China.
Jiangsu Key Laboratory of Sericultural and Animal Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, Jiangsu, China; Key Laboratory of Silkworm and Mulberry Genetic Improvement, Ministry of Agriculture and Rural Affairs, Sericultural Research Institute, Chinese Academy of Agricultural Science, Zhenjiang 212100, Jiangsu, China.
Pestic Biochem Physiol. 2025 Nov;214:106560. doi: 10.1016/j.pestbp.2025.106560. Epub 2025 Jul 10.
The molecular mechanism of baculovirus infection is the basis of baculovirus wide application. Identifying and elucidating the functional genes of virus replication is the focus of research. Eukaryotic initiation factor 4E (eIF4E) is a key component of the translation initiation process to synthesize proteins required for replication. Our pre-transcriptomics data suggest that BmeIF4E is associated with Autographacalifornica nucleopolyhedrovirus (AcMNPV) infection, but its exact role is unknown. The study first clarified, using bioinformatics, that the function of BmeIF4E2 is highly conserved, and expression profile analysis revealed that the high expression of BmeIF4E2 is mainly distributed in the egg stage, 4th instar molting stage, and pupal stage. To clarify the function of BmeIF4E2 in AcMNPV infestation, it was overexpressed and interfered in BmN cells. The increase of virus after overexpression and decrease of virus after interference indicated that BmeIF4E2 could promote AcMNPV infection. To analyze whether BmeIF4E2 is directly involved in promoting AcMNPV infection, His-tag pull down analysis was performed with prokaryotic expression of BmeIF4E2 protein as bait. It was found that BmeIF4E2 could interact directly with vAcMNPV-eGFP. Finally, we validated this in individual silkworms using the inhibitor 4E2RCat and the recombinant virus AcMNPV-BmeIF4E2, and the results were consistent with those obtained in cells. Thus, BmeIF4E2 is a key gene that promotes AcMNPV infection in silkworms via direct interaction. The results are valuable for pest biological control and improving the yield of baculovirus expression systems.
杆状病毒感染的分子机制是杆状病毒广泛应用的基础。鉴定和阐明病毒复制的功能基因是研究的重点。真核生物起始因子4E(eIF4E)是合成复制所需蛋白质的翻译起始过程的关键组成部分。我们的转录组学前期数据表明,BmeIF4E与苜蓿银纹夜蛾核型多角体病毒(AcMNPV)感染有关,但其确切作用尚不清楚。该研究首先利用生物信息学阐明,BmeIF4E2的功能高度保守,表达谱分析表明,BmeIF4E2的高表达主要分布在卵期、4龄蜕皮期和蛹期。为了阐明BmeIF4E2在AcMNPV侵染中的功能,在BmN细胞中对其进行了过表达和干扰。过表达后病毒增加,干扰后病毒减少,表明BmeIF4E2可以促进AcMNPV感染。为了分析BmeIF4E2是否直接参与促进AcMNPV感染,以原核表达的BmeIF4E2蛋白为诱饵进行了His-tag下拉分析。发现BmeIF4E2可以直接与vAcMNPV-eGFP相互作用。最后,我们使用抑制剂4E2RCat和重组病毒AcMNPV-BmeIF4E2在个体家蚕中进行了验证,结果与在细胞中获得的结果一致。因此,BmeIF4E2是通过直接相互作用促进家蚕中AcMNPV感染的关键基因。这些结果对于害虫生物防治和提高杆状病毒表达系统的产量具有重要价值。