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

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.