首页|2群坦布苏病毒NS1蛋白抑制鸭Ⅰ型干扰素产生的机制研究

2群坦布苏病毒NS1蛋白抑制鸭Ⅰ型干扰素产生的机制研究

扫码查看
原文链接
  • 万方数据
  • 维普
[目的]坦布苏病毒(Tembusu virus,TMUV)病是重要的水禽传染病,研究发现TMUV感染鸭早期,2 群TMUV在肝、肾、脑等组织中的病毒拷贝数显著高于 3 群TMUV.研究 2 群TMUV非结构蛋白(Non-structural protein,NS protein)和 3 群TMUV NS蛋白对鸭天然免疫反应是否存在差异.[方法]以 2 群TMUV-JM株和 3 群TMUV-GX株为研究对象,构建两种毒株NS蛋白真核表达质粒,通过双荧光素酶报告系统研究NS蛋白对视黄酸诱导基因蛋白I(Retinoic acid-inducible gene protein I,RIG-I)诱导的β干扰素(β interferon,IFN-β,为I型)启动子活性的影响.构建重组嵌合NS1 蛋白真核表达质粒,通过激光共聚焦、免疫共沉淀、Western Blotting技术研究NS1 与RIG-I信号通路中关键分子的互作区域.利用反向遗传操作系统构建NS1 重组嵌合病毒,研究NS1 在TMUV感染DEF细胞后对IFN-β产生的抑制作用.[结果]TMUV-JM株NS1 能抑制RIG-I诱导的IFN-β启动子活性,而TMUV-GX株NS1 对其无抑制作用.进一步研究发现,TANK结合激酶 1(TANK-binding kinase 1,TBK1)为TMUV-JM NS1 蛋白抑制RIG-I介导的I型IFN表达的作用节点分子,且NS1 蛋白 255～352 aa为发挥抑制作用的功能区域.TMUV-JM NS1与TBK1不存在直接相互作用,是间接降低TBK1磷酸化水平从而降低I型IFN的表达量.[结论]2 群TMUV NS1 具有抑制RIG-I信号通路的活性,并鉴定出其抑制功能的关键活性区域及作用的通路节点分子,明确 2 群TMUV NS1 通过间接抑制TBK1 磷酸化而影响鸭I型IFN产生.
Research on NS1 Proteins of Group 2 Tembusu Virus Inhibiting the Production of Duck Type Ⅰ IFN
[Objective]Tembusu virus(TMUV)disease is an important infectious disease in waterfowl.In previous studies,we found that in the early stages of TMUV infection in ducks,the viral copy number of group 2 TMUV in organs such as the liver,kidney,and brain were significantly higher than those of group 3 TMUV.The study aimed to explore whether group 2 TMUV NS proteins had different effects on the innate immune response of ducks compared with group 3 TMUV.[Method]Taking the group 2 TMUV-JM strain and the group 3 TMUV-GX strain as research subjects,we constructed the eukaryotic expression plasmids of NS proteins of the two strains and compared the effects of the NS proteins on the RIG-I-induced activity of the IFN-β(Type I)promoter by a dual-Luciferase reporter system.With the eukaryotic expression plasmids of the constructed recombinant chimeric NS1 proteins,we investigated the interaction region between NS1 and the key molecules of RIG-I signaling pathway by confocal laser scanning microscopy,co-IP and Western Blotting.By using the reverse genetic system,we constructed NS1 recombinant chimeric TMUV to clarify the inhibitory effect of NS1 on the IFN-β in DEF cells with TMUV infection.[Result]The study results showed that TMUV-JM NS1 could inhibit RIG-I-induced activation of the IFN-β promoter,while TMUV-GX NS1 did not exhibit the inhibitory activity.Further studies revealed that the TMUV-JM NS1 inhibited the expression of type I IFN via targeting TBK1,and the NS1 255-352 aa was the functional region of the inhibitory effect.It was found that TMUV-JM NS1 did not interact with TBK1 directly,but reduced the phosphorylation level of TBK1 indirectly,thereby suppressed the expression of type I IFN.[Conclusion]Based on the study results,we found that the group 2 TMUV NS1 has activity in inhibiting the RIG-I signaling pathway,identified the key activity region and targeted molecule of its inhibitory pathway,and clarified that group 2 TMUV NS1 affected duck type I IFN production by indirectly inhibiting the phosphorylation of TBK1.

Tembusu virusnon-structural proteinRIG-I signaling pathwaytype I interferonTBK1 phosphorylation

黄允真、张俊勤、李林林、董嘉文、向勇、廖明、孙敏华

展开 >

广东省农业科学院动物卫生研究所/农业农村部禽流感等家禽重大疾病防控重点实验室/广东省畜禽疫病防治研究重点实验室,广东 广州 510640

坦布苏病毒 非结构蛋白 RIG-I信号通路 I型干扰素 TBK1磷酸化

国家自然科学基金国家自然科学基金广东省科技计划项目广东省"十四五"农业科技创新十大主攻方向"揭榜挂帅"项目广东省现代农业产业技术体系创新团队项目

32102691319022722021B12120300152022SDZG022023KJ137

2024

10.16768/j.issn.1004-874X.2024.07.014

广东农业科学
广东省农业科学院 华南农业大学

广东农业科学

CSTPCD
影响因子：0.556
ISSN：1004-874X
年,卷(期)：2024.51(7)