Jiangsu Key Laboratory of Sericultural and Animal Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China.
Jiangsu Key Laboratory of Sericultural and Animal Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China; Key Laboratory of Silkworm and Mulberry Genetic Improvement, Ministry of Agriculture and Rural Affairs, Sericultural Scientific Research Center, Chinese Academy of Agricultural Sciences, Zhenjiang 212100, China.
Pestic Biochem Physiol. 2024 Sep;204:106046. doi: 10.1016/j.pestbp.2024.106046. Epub 2024 Jul 25.
Bombyx mori nucleopolyhedrovirus (BmNPV) is a major pathogen that threatens the growth and sustainability of the sericultural industry. Currently, accumulated studies showed that long non-coding RNAs (lncRNAs) play important roles in the genesis and progression of various viruses and host-pathogens interactions. However, the functions and regulatory mechanisms of lncRNAs in insect-virus interaction are still limited. In this study, transcriptome sequencing and ribosome profiling sequencing (Ribo-seq) were performed in the BmNPV-infected midgut and control tissue, and a total of 9 differentially expressed (DE) lncRNAs and 27 small ORFs (sORFs) with micropeptide coding potential were identified. Among them, lncRNA XR_001139971.3 (lnc557) is verified to be significantly up-regulated upon BmNPV infection and may have the potential to encode a small peptide (ORF-674). The subcellular localization experiment showed that lnc557 was expressed in the cytoplasm. Overexpression of lnc557 promotes BmNPV replication and vice versa. By combining RNA pull-down, mass spectrometry, protein truncation and RNA immunoprecipitation (RIP) assays, we confirmed that lnc557 can bind to the RRM-5 domain of BmELAVL1 protein. Subsequently, we found that lnc557 could promote the expression of BmELAVL1 by enhancing the stability of BmELAVL1. Further, enhancing the expression of BmELAVL1 can promote the proliferation of BmNPV, while knockdown shows the opposite effect. Our data suggest that lnc557-mediated BmELAVL1 expression enhancement could play a positive role in BmNPV replication, which will provide a new insight into the molecular mechanism of interaction between Bombyx mori and virus.
家蚕核型多角体病毒（BmNPV）是威胁蚕桑业生长和可持续性的主要病原体。目前，已有大量研究表明，长链非编码 RNA（lncRNA）在各种病毒的发生和发展以及宿主-病原体相互作用中发挥着重要作用。然而，lncRNA 在昆虫-病毒相互作用中的功能和调控机制仍有限。在本研究中，对 BmNPV 感染的中肠和对照组织进行了转录组测序和核糖体谱测序（Ribo-seq），共鉴定出 9 个差异表达（DE）lncRNA 和 27 个具有微小肽编码潜力的小开放阅读框（sORF）。其中，lncRNA XR_001139971.3（lnc557）在 BmNPV 感染后被证实显著上调，可能具有编码小肽（ORF-674）的潜力。亚细胞定位实验表明，lnc557 在细胞质中表达。lnc557 的过表达促进 BmNPV 的复制，反之亦然。通过 RNA 下拉、质谱、蛋白质截断和 RNA 免疫沉淀（RIP）实验相结合，我们证实 lnc557 可以与 BmELAVL1 蛋白的 RRM-5 结构域结合。随后，我们发现 lnc557 通过增强 BmELAVL1 的稳定性来促进 BmELAVL1 的表达。进一步增强 BmELAVL1 的表达可以促进 BmNPV 的增殖，而敲低则显示出相反的效果。我们的数据表明，lnc557 介导的 BmELAVL1 表达增强可能在家蚕与病毒相互作用的复制中发挥积极作用，这将为家蚕与病毒相互作用的分子机制提供新的见解。
