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纳尔逊海湾正呼肠病毒（Nelson bay orthoreovirus,NBV）是一种新兴的具有人畜共患潜能的病毒，可诱发人类呼吸道疾病。然而，对其潜在危险性尚不清楚，因此深入研究NBV复制机制对于理解其致病性具有重要意义。目的本研究旨在探讨宿主蛋白FTH1与NBVσNS蛋白相互作用（互作），阐明宿主蛋白对病毒复制的影响。方法构建真核重组质粒pFLAG-CMV-3-FTH1,Western blot验证其在细胞内的表达。将σNS-HA和FTH1-FLAG两种质粒共转至293T细胞中，利用免疫共沉淀和免疫荧光共聚焦验证σNS和FTH1蛋白互作。以NBV感染L929细胞，RT-qPCR检测FTH1基因在不同感染时间和不同MOI值的相对表达量变化；Western blot检测NBV感染后不同时间点FTH1的蛋白表达水平和不同MOI值NBV感染后48 h FTH1的蛋白表达水平通过灰度值分析比较蛋白表达差异。敲低FTH1基因并感染NBV ,Western blot检测σNS在感染后不同时间点的蛋白表达量变化并通过灰度值分析比较蛋白表达差异。结果 成功构建pFLAG-CMV-3-FTH1重组质粒，表达FTH1蛋白；验证σNS和FTH1蛋白互作；NBV感染促进FTH1在基因与蛋白水平的表达，且该促进作用具有时间依赖性；不同感染复数（MOI分别为0、0.01、0.1、1和5）的NBV感染均促进FTH1的表达，且在MOI 5的条件下FTH1蛋白表达量升高最明显；敲低FTH1基因，σNS表达量减低。结论 验证FTH1与σNS蛋白发生互作。FTH1通过与σNS相互作用，促进NBV的复制。研究结果为深入分析NBV复制进程中宿主蛋白与病毒蛋白的相互作用机制奠定了基础，也为探索NBV感染的潜在治疗靶点提供了重要思路。
Abstract：Nelson Bay orthoreovirus(Nelson Bay orthoreovirus, NBV) is an emerging virus with zoonotic potential that can cause respiratory disease in humans. However, its potential risk remains poorly understood. Therefore, elucidating the replication mechanism of NBV is essential for understanding its pathogenicity. Objective To investigate the interaction between the host protein FTH1 and the NBV σNS protein and to clarify the role of this host protein in viral replication. Methods The eukaryotic recombinant plasmid pFLAGCMV-3-FTH1 was constructed, and its expression in cells was verified by Western blotting. Plasmids encoding σNS-HA and FTH1-FLAG were co-transfected into 293T cells, and the interaction between σNS and FTH1 proteins was examined using co-immunoprecipitation and immunofluorescence confocal microscopy. L929 cells were infected with NBV, and the relative expression levels of the FTH1 gene at different time points postinfection and under different multiplicities of infection(MOIs) were measured by RT-qPCR. Western blotting was used to detect FTH1 protein expression at different time points after NBV infection and at 48 h postinfection under different MOI conditions, and differences in protein expression were analyzed by densitometry. After knockdown of the FTH1 gene followed by NBV infection, σNS protein expression at different time points post-infection was detected by Western blotting and analyzed by densitometry. Results The recombinant plasmid pFLAG-CMV-3-FTH1 was successfully constructed and expressed the FTH1 protein. The interaction between σNS and FTH1 proteins was confirmed. NBV infection promoted FTH1 expression at both the mRNA and protein levels in a time-dependent manner. NBV infection at different MOIs(0, 0.01, 0.1, 1, and 5) increased FTH1 expression, with the most pronounced upregulation observed at an MOI of 5. Knockdown of FTH1 resulted in reduced σNS protein expression. Conclusion FTH1 interacts with the NBV σNS protein and promotes NBV replication through this interaction. These findings provide a basis for further investigation of host– virus protein interactions during NBV replication and offer new insights into potential therapeutic targets for NBV infection.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.250410
中图分类号:R373
引用信息:
[1]路佩嘉,钟萍,曹珂铨,等.宿主蛋白FTH1与纳尔逊海湾正呼肠病毒σNS蛋白互作并促进病毒复制[J].病毒学报,2026,42(03):823-832.DOI:10.13242/j.cnki.bingduxuebao.250410.
2026-03-25
2026-03-25
2026-03-25