摘要
家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)作为杆状病毒科ɑ型杆状病毒属的代表性成员,其有效感染依赖于病毒核衣壳穿越宿主细胞核膜进入细胞核。然而,病毒入核过程中是否会破坏宿主核膜结构,目前仍缺乏系统性的分析。在本研究中,通过RNA干扰技术靶向沉默家蚕卵巢细胞(BmN)中的核纤层蛋白LamC和Dm0的表达,并利用70 kD FITC标记葡聚糖(FITC-Dextran)作为大分子通透性探针,对病毒感染过程中细胞核膜完整性进行分析。激光共聚焦显微观察显示,FITC-Dextran在健康或BmNPV感染的细胞中均不能进入细胞核,而在核纤层蛋白敲低的细胞中,FITC-Dextran能进入细胞核中,表明核纤层干扰导致核膜裂解进而使得FITC-Dextran可以入核,而病毒感染并没有发生核膜裂解现象。通过透射电镜进一步分析证实,病毒感染过程中核膜保持结构完整。以上结果表明,BmNPV核衣壳入核不依赖核膜裂解而是通过核孔复合体介导。本研究明确了BmNPV核输入机制的结构基础,为揭示病毒宿主特异性感染机制及开发靶向核转运过程中抗病毒策略提供信息。
Bombyx mori nucleopolyhedrovirus(BmNPV),a representative member of the genus Alphabaculovirus in the family Baculoviridae.It’s effective infection relies on the transport of its nucleocapsid across the host nuclear envelope to enter the nuoleus.However,whether this nuclear entry process involves structural disruption of the nuclear envelope still lack systematic analysis.In this study,RNA interference was employed to knock down the expression of nuclear lamia proteins LamC and Dm0 in BmN cells.Using FITC-labeled dextran(FITC-Dextran)with molecular weight about 70 kD as a macromolecular permeability probe,we analyzed the integrity of the nuclear envelope during BmNPV infection.Laser confocal microscopy observation revealed that FITC-Dextran was excluded from the nucleus in both healthy and BmNPV-infected cells,whereas it entered the nucleus in cells with lamins knockdown,indicating that disruption of nuclear lamins caused lysis of nuclear envelope,allowing nuclear entry of FITC-Dextran.In contrast,BmNPV infection did not induce nuclear envelope lysis.Analysis by transmission electron microscopy further confirmed that the nuclear envelope remained structurally intact during viral infection.Collectively,these results demonstrate that BmNPV nucleocapsid enters the nucleus via the nuclear pore complex,independent of nuclear envelope lysis.This study elucidates the structural basis of BmNPV nuclear import,providing insights into the mechanism of host-specific viral infection and informing the development of antiviral strategies targeting nuclear transport.
作者
田海珏
尹康平
张天然
黄金山
郝碧芳
Tian Haijue;Yin Kangping;Zhang Tianran;Huang Jinshan;Hao Bifang(School of Biotechnology,Jiangsu University of Science and Technology,Zhenjiang Jiangsu 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 Jiangsu 212100,China)
出处
《蚕业科学》
北大核心
2025年第5期437-443,共7页
ACTA SERICOLOGICA SINICA
基金
国家自然科学基金项目(32172796)。
关键词
家蚕核型多角体病毒
核衣壳
核输入
核膜
Bombyx mori nucleopolyhedrovirus
Nucleocapsid
Nuclear import
Nuclear envelope
