宿主蛋白RPSA在非洲猪瘟病毒复制过程中的作用研究  

The role of host protein RPSA in the replication of African swine fever virus

作　　者：邵程祥 张忠辉 孙华林 赵亚茹 杨吉飞[2] 关贵全[2] 殷宏[2,3] 武军元 牛庆丽[2] NIU Qingli;SHAO Chengxiang;ZHANG Zhonghui;SUN Hualin;ZHAO Yaru;YANG Jifei;GUAN Guiquan;YIN Hong;WU Junyuan;NIU Qingli(College of Animal Science and Technology,Tarim University,Xinjiang Aral 843300,China;State Key Laboratory for Animal Disease Control and Prevention/African Swine Fever Regional Laboratory of China(Lanzhou)/College of Veterinary Medicine and Biosafety of Lanzhou University/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonosis,Yangzhou 225009,China)

机构地区：[1]塔里木大学动物科学与技术学院,新疆阿拉尔843300 [2]中国农业科学院兰州兽医研究所兰州大学动物医学与生物安全学院动物疫病防控全国重点实验室国家非洲猪瘟区域实验室,甘肃兰州730046 [3]江苏高校动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《中国兽医科学》2024年第9期1149-1156,共8页Chinese Veterinary Science

基　　金：国家自然科学基金面上项目(32072830);国家重点研发计划项目(2021YFD1800101);农业科技创新工程项目(ASTIP,CAAS-ASTIP-2016-LVRI);甘肃省科技重大专项(22ZD6NA001);甘肃省基础研究创新群体项目(22JR5RA024);甘肃省优秀博士生项目(22JR5RA025)

摘　　要：本研究旨在探究宿主RPSA蛋白对非洲猪瘟病毒(African swine fever virus,ASFV)复制的影响,以及病毒蛋白B602L与RPSA之间的相互作用。利用RPSA重组质粒pcDNA3.1-Flag-RPSA转染293T细胞,通过免疫荧光检测B602L和RPSA蛋白的亚细胞定位,通过免疫共沉淀(Co-IP)验证它们之间的相互关系。通过荧光定量PCR和Western-blot分析ASFV感染IPAM细胞在不同时间点RPSA的表达差异。此外,通过在IPAM细胞中过表达或干扰RPSA的表达,分析RPSA对ASFV复制的影响。研究结果表明,ASFV感染IPAM细胞后,RPSA蛋白的表达随时间延长而逐渐增加,表明RPSA蛋白可能在病毒感染中发挥作用。通过过表达RPSA可抑制病毒复制,而干扰RPSA则导致病毒复制增强,进一步表明RPSA在ASFV复制过程中发挥重要作用。最后,Co-IP和亚细胞定位结果证明病毒蛋白B602L与宿主蛋白RPSA之间存在相互作用。综上所述,本研究揭示了宿主蛋白RPSA在ASFV复制过程中的作用,为进一步研究病毒与宿主之间的相互作用机制奠定了基础,也有助于深入了解ASFV的感染机制为疾病控制提供新的线索。This study aimed to investigate the impact of host protein RPSA the replication of African swine fever virus(ASFV)and the interaction between viral B602L protein and RPSA.The recombinant RPSA plasmid pcDNA3.1-Flag-RPSA was constructed and transfected into HET 293 cells,and the subcellular localization of B602L and RPSA proteins was examined through immunofluorescence assay(IFA),as well as their interaction was validated through immunoprecipitation(Co-IP).Additionally,the expressed differences of RPSA protein during ASFV infected different time points in IPAM cells were analyzed through quantitative real-time PCR(qPCR)and Western blot.Furthermore,the impact of RPSA on the ASFV replication process was analyzed by over-expression and interfering of RPSA expression in IPAM cells.The results showed that the expression of RPSA protein gradually increased over time following ASFV infection in IPAM cell Over-expression of RPSA could inhibit viral replication,while interference of RPSA leads to enhance of viral replication.Moreover,the results of Co-IP and subcellular localization were also confirmed that the interaction between viral B602L protein and host RPSA protein.In summary,this study revealed the critical role of host RPSA protein in the ASFV replication process,providing an important theoretical basis for further mechanism research.These findings contribute to a deeper understanding of the infection mechanism of ASFV infection and provide new clues for disease control and treatment.

关 键 词：非洲猪瘟病毒 B602L RPSA 相互作用 

分 类 号：S852.659.1[农业科学—基础兽医学]