EV71病毒诱导细胞应激颗粒形成的初步研究

Preliminary Study on the Formation of Cellular Stress Granules Induced by EV71 Virus

肠道病毒71型(Enterovirus 71,EV71)属于小RNA病毒科,肠道病毒属,肠道病毒A型的成员,是引起婴幼儿手足口病(Hand foot and mouth disease,HFMD)的一种重要病原病毒。但截至目前,EV71病毒与宿主细胞相互作用的机制仍不清楚。为了研究EV71病毒诱导应激颗粒形成的分子机制,研究首先构建了真核表达载体pDsRed2-G3BP1,随后构建了稳定表达融合蛋白RFP-G3BP1蛋白的HeLa细胞系(HeLaRFP-G3BP1),并应用Western Blot和免疫荧光技术证实了G3BP1融合蛋白的正确表达。EV71病毒感染HeLaRFP-G3BP1后,超过40%感染细胞的胞质中出现了应激颗粒。此外,应用微管特异性抑制剂破坏细胞微管完整性之后,显著抑制应激颗粒的迁移聚集。研究证明了EV71病毒诱导细胞应激颗粒的初步形成并且应激颗粒的迁移聚集依赖于微管的完整性。

Enterovirus 71(Enterovirus 71,EV71)is A member of the small RNA virus family,the genus enterovirus,and the type A of enterovirus,and is an important pathogenic virus causing hand-foot-mouth disease(Hand foot and mouth disease,HFMD)in infants.However,the main reason is that the mechanism for the interaction of EV71 with host cells is still unclear.To investigate the molecular mechanism of EV71 induced the initial formation of cellular SGs.In this study,we have firstly constructed the eukaryotic expression vector pDsRed2-G3BP1 and constructed a HeLa cell line(HeLaRFP-G3BP1)stably expressing the fusion protein RFP-G3BP1.Western blot and immunofluorescence methods were used to confirm the correct expression of the G3BP1 fusion protein.HeLaRFP-G3BP1 containing(SGs)in their cytoplasm were more than 40%of total cells during EV71 infection.In addition,after the microtubule specific inhibitor was used to destroy the integrity of microtubule,the migration and aggregation of SGs were significantly inhibited.This study demonstrates that EV71 induces the initial formation of SGs and that migration of SGs is dependent on the integrity of the microtubules.