PIAS1对PRRSV N蛋白SUMO化修饰及病毒复制的影响

发布时间：2018-04-19 06:01

本文选题：PRRSV + N蛋白　；参考：《微生物学通报》2017年12期

【摘要】：【目的】猪繁殖与呼吸综合征病毒(PRRSV)是一种危害全球养猪业的重要病原。SUMO(Small ubiquitin-like modifier)化修饰作为一种可逆的翻译后修饰在调节病毒复制方面发挥着重要功能。PIAS1(Protein inhibitor of activated STAT1)是SUMO E3连接酶PIAS家族的一员,可以促进靶蛋白的SUMO化修饰,进而影响靶蛋白的功能,参与基因转录调控过程。探究PIAS1与PRRSV N蛋白相互作用的机制及其对N蛋白SUMO化修饰和病毒复制的影响,为进一步阐明PRRSV复制调控和致病的分子机制提供科学依据。【方法】利用酵母回复杂交、免疫共沉淀和激光共聚焦技术验证N蛋白与PIAS1的相互作用;以递增剂量外源性转染PIAS1观察其是否介导N蛋白SUMO化修饰;采用RNA干扰和慢病毒转导技术测定PIAS1对PRRSV复制的影响。【结果】PIAS1能与N蛋白相互作用,而且两者主要共定位于胞浆中;外源转染PIAS1并未增加N蛋白SUMO化修饰水平;在MARC-145细胞中,PIAS1的表达有利于PRRSV的复制。【结论】PIAS1可促进PRRSV的复制。
[Abstract]:[objective] Porcine Reproductive and Respiratory Syndrome virus (PRRSVV) is a major pathogen of swine industry worldwide. Sumo small ubiquitin-like modifier, as a reversible posttranslational modifier, plays an important role in regulating viral replication. PIAS1 protein inhibitor of activated STAT1)Is a member of the SUMO E3 ligase PIAS family,It can promote the SUMO modification of the target protein, and then affect the function of the target protein, and participate in the process of gene transcription regulation.To explore the mechanism of interaction between PIAS1 and PRRSV N protein and its effect on SUMO modification of N protein and viral replication, to provide scientific basis for further elucidating the molecular mechanism of PRRSV replication and pathogenesis. [methods] yeast reverse hybridization was used.Immunoprecipitation and laser confocal technique were used to verify the interaction between N protein and PIAS1, and to observe whether N protein SUMO modification was mediated by exogenous transfection of PIAS1 with increasing dose.RNA interference and lentivirus transduction techniques were used to determine the effect of PIAS1 on PRRSV replication. [results] PIAS1 could interact with N protein, and both of them were mainly located in cytoplasm, and exogenous PIAS1 transfection did not increase the level of SUMO modification of N protein.The expression of PIAS1 in MARC-145 cells is beneficial to the replication of PRRSV. [conclusion] PIAS1 can promote the replication of PRRSV.
【作者单位】：中国农业大学动物医学院农业部动物流行病学重点实验室;
【基金】：国家自然科学基金面上项目(No.31572549)~~
【分类号】：S852.651

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