羊传染性脓疱病毒ORF120蛋白以剂量依赖性方式抑制宿主细胞非POU结构域八聚体结合蛋白(NONO)的表达

Orf virus ORF120 protein inhibits the expression of non-POU domain-containing octamer-binding protein(NONO)in a dose-dependent manner

羊传染性脓疱病毒(ORFV)是重要的人兽共患病病原,能够通过劫持宿主蛋白等多种策略逃避机体的抗病毒免疫反应,导致重复感染的发生。非POU结构域八聚体结合蛋白(NONO)是一种多功能核蛋白,参与mRNA剪切、DNA损伤修复、肿瘤发生以及炎症反应等多种生物学过程。本研究首先利用酵母双杂交、免疫共沉淀(Co-IP)等技术证实ORF120蛋白与宿主蛋白NONO存在相互作用。借助于激光共聚焦显微镜观察病毒ORF120蛋白和宿主NONO蛋白的定位分布,发现ORF120蛋白与NONO蛋白在细胞核内存在共定位,进一步证实ORF120蛋白和NONO蛋白存在互作。此外,利用Western blot方法证实ORF120蛋白和NONO蛋白之间存在相互抑制作用,且呈剂量依赖性。上述结果表明,ORF120蛋白与宿主NONO蛋白存在剂量依赖性的相互抑制作用,该结果将为更好地理解ORFV免疫逃逸分子机制及开发新的抗病毒手段提供重要依据。

Sheep infectious pustular virus(ORFV)is an important zoonotic pathogen,which can escape the antiviral immune response of the body by hijacking the host protein and other strategies,leading to repeated infection.Non-POU domain octamer binding protein(NONO)is a multifunctional nuclear protein,which participates in many biological processes such as mRNA splicing,DNA damage repair,tumorigenesis and inflammatory response.In this study,the interaction between ORF120protein and host protein NONO was firstly confirmed by yeast two-hybrid and immunoprecipitation(Co-IP).The localization and distribution of virus ORF120protein and host NONO protein were observed by laser confocal microscopy.It was found that ORF120protein and NONO protein co-located in the nucleus,further confirming the interaction between ORF120protein and NONO protein.In addition,Western blot method was used to confirm the mutual inhibition between ORF120protein and NONO protein in a dose-dependent manner.The above results show that ORF120protein and host NONO protein have a dose-dependent interaction,which will provide an important basis for better understanding the molecular mechanism of ORFV immune escape and developing new antiviral methods.