| 金红梅，袁春颖，邹德馨，侯春生，马鸣潇，邓炎春,2025,西方蜜蜂NPC2基因的生物信息学分析及其在IAPV感染过程中的功能鉴定[J].环境昆虫学报,(4):1238-1249 |
| 西方蜜蜂NPC2基因的生物信息学分析及其在IAPV感染过程中的功能鉴定 |
| Bioinformatic analysis and functional characterization of the NPC2 gene in Apis mellifera during IAPV infection |
| |
| DOI： |
| 中文关键词: 西方蜜蜂 转运胆固醇蛋白 分子特性 表达特征 以色列急性麻痹病毒 |
| 英文关键词:Apis mellifera NPC intracellular cholesterol transporter 2 Molecular characteristics Expression profile IAPV |
| 基金项目:国家自然科学基金青年基金项目（32300418）；湖南省科技领军人才项目（2024RC1069） |
|
| 摘要点击次数: 478 |
| 全文下载次数: 420 |
| 中文摘要: |
| 本研究旨在探究西方蜜蜂Apis mellifera细胞内胆固醇转运蛋白2（NPC intracellular cholesterol transporter 2，AmNPC2）的分子特性及其在以色列急性麻痹病毒（Israeli acute paralysis virus，IAPV）感染过程中的潜在调控作用。通过综合运用生物信息学分析、实时荧光定量PCR（RT-qPCR）和RNA干扰（RNAi）等技术手段，研究了AmNPC2的分子特征、组织表达谱及其对IAPV入侵的调控功能。结果表明，AmNPC2基因CDS区域长度为465 bp，对应开放阅读框（ORF）编码154个氨基酸。AmNPC2蛋白的相对分子量约为16.8 kDa，含14个磷酸化位点，具有跨膜结构域；含信号肽，剪切位点位于17~18号氨基酸之间，主要定位于细胞外；系统进化分析显示AmNPC2与东方蜜蜂Apis cerana的NPC2聚为一支；二级结构预测显示，AmNPC2包含83个无规则卷曲、26个α-螺旋、39条延伸链和6个β-转角。RT-qPCR结果显示，AmNPC2在自然感染IAPV的工蜂中的8个检测组织中均有表达，其中在蜜囊和气管中表达量最高，而在中肠中表达相对较低；值得注意的是，IAPV自然感染主要也发生在蜜囊和气管组织。通过实验感染模型分析发现，IAPV感染后，蜜蜂AmNPC2基因的转录水平显著上调。通过RNA干扰技术特异性敲降AmNPC2基因的表达后，定量分析表明IAPV病毒的复制效率出现了显著抑制，其病毒载量下降了6倍以上。本研究结果表明AmNPC2可能通过调控宿主脂质代谢参与IAPV的感染过程，该研究首次揭示了NPC2基因在蜜蜂病毒过程中的潜在作用，为深入理解蜜蜂-病毒互作机制提供了新的理论依据。 |
| 英文摘要: |
| This study aimed to investigate the molecular characteristics of NPC intracellular cholesterol transporter 2 (AmNPC2) in Apis mellifera and its potential regulatory role during Israeli acute paralysis virus (IAPV) infection. Through integrated approaches including bioinformatics analysis, quantitative real-time PCR (qRT-PCR), and RNA interference (RNAi), we examined the molecular features, tissue expression profile, and regulatory function of AmNPC2 in IAPV replication. Bioinformatics analysis revealed that the AmNPC2 gene contains a 465bp coding sequence (CDS) encoding a 154-amino acid protein with a predicted molecular weight of 16.8 kDa. Structural analysis revealed 14 phosphorylation sites, a transmembrane domain, and an N-terminal signal peptide (cleavage site between residues 17-18), indicating extracellular localization. Phylogenetic analysis showed that AmNPC2 clusters with NPC2 from Apis cerana, and secondary structure prediction identified 26 α-helices, 39 β-strands, 6 β-turns, and 83 random coils. RT-qPCR analysis demonstrated ubiquitous AmNPC2 expression across eight examined tissues in worker bees, with highest levels in the honey sac and trachea-tissues that also showed predominant IAPV tropism during natural infection. Experimental infection studies demonstrated that IAPV infection induces a significant upregulation of AmNPC2 gene expression in the host. RNAi-mediated knockdown of AmNPC2 led to more than 6-fold reduction in viral loads, suggesting that AmNPC2 facilitates IAPV infection, potentially through modulation of host lipid metabolism. Our results demonstrated that AmNPC2 may participate in IAPV infection by regulating host lipid metabolism. This study provided the first evidence for the potential role of NPC2 genes in honeybee viral pathogenesis, offering new theoretical insights into bee-virus interaction mechanisms. |
| 查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
