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目的 非洲猪瘟（African swine fever, ASF）是一种由非洲猪瘟病毒（African swine fever virus, ASFV）引起的高度致死性传染病，可导致家猪和野猪出现严重出血性症状及高死亡率。环指蛋白5(Ring finger protein 5, RNF5)是一种E3泛素连接酶，参与泛素介导的蛋白降解及炎症信号通路调控。本研究旨在探究ASFV感染过程中RNF5的作用及其介导的宿主蛋白互作网络。方法 首先在体外评估RNF5过表达对ASFV复制的影响；随后采用免疫共沉淀-质谱（Co-immunoprecipitation-mass spectrometry, Co-IP-MS）筛选RNF5相互作用蛋白，并对差异富集蛋白的生物学功能进行分析；最后在ASFV感染条件下，通过免疫共沉淀实验对候选互作蛋白进行验证。结果 RNF5过表达显著抑制ASFV复制，表明其可能具有抗病毒作用。Co-IP-MS分析鉴定出多种与RNF5存在潜在互作的候选蛋白。功能分析结果表明，这些蛋白主要参与转录与RNA加工、代谢及信号转导等生物学过程。免疫共沉淀实验进一步证实RNF5可与DEAD-box RNA解旋酶3 X连锁蛋白（DEAD-box helicase 3 X-linked, DDX3X）和B细胞淋巴瘤-2(B-cell lymphoma-2, BCL-2)发生特异性相互作用。在ASFV感染条件下，DDX3X表达水平显著升高，而BCL-2表达无明显变化，但二者与RNF5的相互作用均显著增强。结论 本研究证实RNF5对ASFV复制具有抑制作用，并进一步解析了其潜在的宿主蛋白互作网络。研究结果为理解ASFV如何调控宿主蛋白互作网络，从而影响病毒复制及宿主细胞反应提供了新的理论依据。
Abstract：Objective African swine fever(ASF) is a highly lethal infectious disease caused by African swine fever virus(ASFV), resulting in severe hemorrhagic symptoms and high mortality in domestic pigs and wild boars. Ring finger protein 5(RNF5) is an E3 ubiquitin ligase involved in ubiquitin-mediated protein degradation and inflammatory signaling pathways. This study aimed to investigate the role of RNF5 and its mediated host protein interaction network during ASFV infection. Methods The effect of RNF5 overexpression on ASFV replication was first evaluated in vitro. Subsequently, co-immunoprecipitation coupled with mass spectrometry(Co-IP-MS) was performed to identify RNF5-interacting proteins, followed by functional analysis of differentially enriched proteins. Finally, candidate interacting proteins were further validated by coimmunoprecipitation assays under ASFV infection conditions. Results RNF5 overexpression significantly inhibited ASFV replication, indicating its potential antiviral activity. Co-IP-MS analysis identified multiple candidate proteins potentially interacting with RNF5. Functional analysis indicated that these proteins were mainly involved in biological processes including transcription and RNA processing, metabolism, and signal transduction. Co-immunoprecipitation assays further confirmed that RNF5 specifically interacted with DEADbox helicase 3 X-linked(DDX3X) and B-cell lymphoma-2(BCL-2). Under ASFV infection conditions, the expression level of DDX3X was significantly upregulated, whereas BCL-2 expression showed no obvious change; however, the interactions of both proteins with RNF5 were significantly enhanced following ASFV infection. Conclusion This study confirms the inhibitory effect of RNF5 on ASFV replication and further characterizes its potential host interaction network. The findings provide new insights into how ASFV may modulate host protein interaction networks to influence viral replication and host cellular responses.
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基本信息:
中图分类号:S858.28
引用信息:
[1]刘颖,孙华林,陈奕康,等.环指蛋白5调控非洲猪瘟病毒感染的关键互作蛋白的筛选与鉴定[J].病毒学报().
基金信息:
国家自然科学基金项目（项目号：U24A20450）,题目：蜱传播非洲猪瘟病毒的分子机制研究; 甘肃省自然科学基金重点项目（项目号：25JRRA436）,题目：蜱Rho/ROCK信号通路调控非洲猪瘟病毒复制的分子机制及抗病毒制剂研究; 甘肃省青年科技基金（项目号：26JRRA385）,题目：拉合尔钝缘蜱影响ASFV传播的唾液腺分泌蛋白鉴定及其分子机制研究
2026-06-02
2026-06-02
2026-06-02