中文摘要
猪繁殖与呼吸综合征病毒(Porcine reproductive and respiratory syndrome virus,PRRSV)可引起宿主免疫抑制和持续性感染,至今仍缺乏有效的防治措施。P-body是细胞内降解、储存mRNA的重要场所,我们在前期的研究中发现P-body重要组件LSm14A在PRRSV感染过程中mRNA和蛋白水平均发生了变化。PRRSV全病毒与病毒的非结构蛋白2(NSP2)均表现出对LSm14A天然免疫活性的拮抗作用。但是,LSm14A等P-body组分是否在PRRSV持续感染中的作用以及与PRRSV的相互作用机理尚不清楚。本项目拟在此基础上从病毒与宿主相互作用入手,通过激光共聚焦定位、质谱分析等方法,明确影响PRRSV复制的P-body组件,研究LSm14A及其它P-body组件蛋白在PRRSV感染与复制过程中的作用机制,最终为PRSSV防控提供新的策略。
英文摘要
Porcine Reproductive and Respiratory Syndrome (PRRS), caused by Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), a RNA virus, is characterized by immunosuppression and persistent infection. PRRS brings about significant economic losses in the pig industry, yet effective therapeutic methods have not been elucidated. P-body is an important site for the degradation and storage of intracellular mRNA. In our previous studies, we expressed LSm14A (RAP55/SCD6), an important component of P-body in vitro, and found that expression of LSm14A was compromised during PRRSV infection. Both virion and non-structural protein 2 (NSP2) of PRRSV showed antagonistic effect on LSm14A mediated innate immune activity. However, the roles of the P-body components (e.g. LSm14A) in PRRSV persistent infection and their interactions to PRRSV structural or nonstructural proteins, remain unclear. Therefore, the PRRSV replication and persist infection mechanisms regulated by P-body components will be explored by confocal laser imaging scanning, co-immunoprecipitation, immunoblotting and mass spectrometry approaches. The colocalization of LSm14A and viral proteins will be characterized in PRRSV infected T cells to reveal mRNA diminish and accumulation kinetics. Then, the localization pattern of P-body components will be investigated on the viral replication sites. This will provide reliable information to clarity potential role of RNA processing machinery in PRRSV infectious pathogenesis for virus replication, disease prevention and control.??
