基于Src磷酸化特异性调控RLRs信号通路揭示鸭抗病毒遗传特性

鸭体内存在RIG-I是鸭与鸡(缺失RIG-I)对禽流感产生先天免疫反应差异主要原因，但鸭如何依赖RIG-I特异性调节病毒清除机制尚不清楚。项目前期高通量测序发现，酪氨酸蛋白激酶Src可特异性调节鸭抗病毒免疫应答，且TRIM25（E3泛素化连接酶）磷酸化也参与这一过程。据此提出鸭Src可能磷酸化TRIM25介导RIG-I泛素化特异性调控RLRs通路的假说。本项目通过检测过表达/敲低Src和不同Src抑制剂对报告基因(NF-κB, IRF3和IFN-β)活性的影响，确定鸭Src对RLRs信号通路中的抗病毒免疫应答作用；综合运用pull-down、IP结合定点突变等技术，重点探索Src磷酸化TRIM25介导RIG-I泛素化活化RLRs通路分子机制。研究将从表观遗传学层面填补鸭特有免疫调节机制的科学数据，这不仅有助于揭开鸭特有的病毒清除机制神秘面纱，还可为禽流感防控提供新的解决方案。

The RIG-I gene is presence in ducks, which is the main reason for the innate immune responding to avian influenza compared with chickens (without RIG-I). However, the mechanism that how ducks rely on RIG-I to regulate the virus clearance is unclear. High-throughput sequencing found that tyrosine protein kinase Src could specially regulate duck antiviral immune response, and TRIM25(E3 ubiquitination ligase) phosphorylation was also involved in this process. Based on this, it is suggested that duck Src may phosphorylate TRIM25 to mediate RIG-I ubiquitination and specifically regulate RLRs pathway. In this project, overexpression/knockdown of Src and different Src inhibitors were used to detect the activity of NF-κB, IRF3 and IFN-β to determine the antiviral immune response of duck Src on the RLRs signaling pathway. Furthermore, pull-down technology, combined with immunoprecipitation and site-directed mutation was used to explore the molecular mechanism that Src phosphorylated TRIM25-mediated RIG-I ubiquitination activation of RLRs pathway. The research will supplement the scientific data of specific immune regulation mechanism from the epigenetics in duck, which will not only help to reveal the mystery of specific virus clearance mechanism, but also provide a new solution for the prevention and control of avian influenza.