鸡G3BP1在新城疫病毒感染诱导应激颗粒形成过程中的作用

发布时间：2019-01-30 19:30

【摘要】：为掌握鸡细胞中应激颗粒(stress granules,SGs)形成机制,以及鸡GTP酶激活蛋白SH3功能区结合蛋白1(G3BP1)在新城疫病毒(NDV)感染诱导SG形成过程中的作用,用NDV感染HeLa细胞,免疫荧光试验检测内源性G3BP1的定位,外源转染鸡G3BP1和各分段结构域,以不同应激处理之后观察与内源性SG标志物共定位现象,最后转染G3BP1之后感染NDV,以Western blot和TCID50检测病毒复制情况。结果显示:NDV感染能够诱导内源性G3BP1聚集形成SG,将鸡G3BP1分别转染至HeLa、DF-1和CEF后,以不同应激处理之后均能在细胞中观察到SG。将鸡G3BP1不同结构域分别转染HeLa细胞,以氧化应激和NDV分别处理,结果显示G3BP1的RNA结合结构域对聚集至关重要,而NTF2样结构域不能被招募到SGs中,并且抑制原有SGs的形成。最后证实G3BP1过表达能够增强病毒复制,说明SG在NDV复制过程中发挥促进作用。综合以上结果表明:鸡G3BP1在鸡SG形成过程中至关重要,鸡SG能够促进NDV复制,这为进一步研究NDV和禽源细胞互作奠定了基础。
[Abstract]:In order to understand the mechanism of stress granules (stress granules,SGs) formation in chicken cells and the role of SH3 binding protein 1 (G3BP1) in SG formation induced by Newcastle disease virus (NDV), NDV was used to infect HeLa cells. The localization of endogenous G3BP1 was detected by immunofluorescence assay. After exogenous transfection of chicken G3BP1 and its segmental domains, co-localization with endogenous SG markers was observed after different stress treatments. Finally, NDV, was infected after transfection of G3BP1. Western blot and TCID50 were used to detect viral replication. The results showed that NDV infection could induce endogenous G3BP1 aggregation to form SG, transfection of chicken G3BP1 into HeLa,DF-1 and CEF, and SG. could be observed in cells after different stress treatments. Different domains of chicken G3BP1 were transfected into HeLa cells and treated with oxidative stress and NDV respectively. The results showed that the RNA binding domain of G3BP1 was very important for aggregation, but NTF2 like domain could not be recruited into SGs and inhibited the formation of SGs. Finally, it was confirmed that G3BP1 overexpression could enhance viral replication, indicating that SG played a role in NDV replication. The above results show that chicken G3BP1 is very important in the formation of chicken SG, and chicken SG can promote NDV replication, which lays a foundation for further study on the interaction between NDV and avian origin cells.
【作者单位】：山东农业大学动物科技学院;中国农业科学院上海兽医研究所;吉林农业大学动物科学技术学院;
【基金】：国家自然科学基金重点项目(31530074);国家自然科学基金面上项目(31372421);国家自然科学基金青年基金(31400144)
【分类号】：S852.65

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