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痘病毒编码多种能够操控宿主先天免疫反应的蛋白。研究表明，鼠痘病毒编码的锚蛋白通过与Skp1互作抑制NF-κB通路。山羊痘病毒编码的锚蛋白ORF141.2与鼠痘病毒编码的锚蛋白存在类似的结构，其对NF-κB信号通路的调控作用还未见报道。为了明确ORF141.2对NF-κB信号通路的调控作用，本研究采用双荧光素酶报告系统检测了ORF141.2对NF-κB转录活性的影响；通过Western bloting技术和间接免疫荧光试验检测了ORF141.2对NF-κB通路关键因子的影响及ORF141.2与Skp1的共定位情况；最后采用Co-IP技术验证了ORF141.2与Skp1是否互作。结果表明，锚蛋白ORF141.2可以抑制NF-κB的转录活性；锚蛋白ORF141.2不影响IKKα和IκBα的磷酸化，但阻止p-IκBα的降解和p65的入核；锚蛋白ORF141.2与Skp1共定位于细胞质中，存在互作关系。锚蛋白ORF141.2通过与Skp1互作调控p-IκBα的降解和p65的入核，从而调控NF-κB信号通路。
Abstract：Poxviruses encode a range of proteins that modulate the host innate immune response. Previous studies have shown that the ankyrin protein encoded by ectromelia virus suppresses the NF-κB signaling pathway through interaction with Skp1. The goatpox virus(GTPV) encodes a structurally similar ankyrin protein, ORF141.2; however, its role in NF-κB regulation remains uncharacterized. In this study, a dual-luciferase reporter assay was used to evaluate the effect of ORF141.2 on NF-κB transcriptional activity. Western blotting and indirect immunofluorescence assays were employed to analyze the impact of ORF141.2 on key NF-κB pathway components and its subcellular co-localization with Skp1. Co-immunoprecipitation(Co-IP) assays were further conducted to verify the physical interaction between ORF141.2 and Skp1. The results revealed that ORF141.2 significantly inhibits NF-κB transcriptional activity. While it does not affect the phosphorylation of IKKα and IκBα, ORF141.2 blocks the degradation of phosphorylated IκBα(p-IκBα) and impedes the nuclear translocation of p65. Moreover, ORF141.2 co-localizes with Skp1 in the cytoplasm and directly interacts with it.These findings suggest that GTPV ankyrin protein ORF141.2 modulates the NF-κB signaling pathway by interacting with Skp1, thereby interfering with p-IκBα degradation and preventing p65 nuclear translocation.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.240325
中图分类号:S852.65
引用信息:
[1]杨云凤,常慧慧,冯秋媛,等.山羊痘病毒锚蛋白ORF141.2对NF-κB信号通路的调控作用研究[J].病毒学报,2025,41(04):1045-1055.DOI:10.13242/j.cnki.bingduxuebao.240325.
基金信息:
国家自然科学基金项目（项目号：32260874）,题目：羊痘病毒RINGfinger蛋白对NF-κB通路的调控作用研究~~
2025-07-11
2025-07-11
2025-07-11