病毒感染的进展涉及许多转录调控事件。鉴定新合成的转录本有助于我们了解病毒的复制机制和发病机理。这里,我们利用一种称为硫醇(SH)连接烷基化的代谢RNA标记方法的时间分辨技术对RNA进行代谢测序(SLAM-seq),以差异阐明BHK21细胞系响应人冠状病毒OC43(HCoV-OC43)感染时稳态和新合成的RNA水平.我们的结果表明,响应于HCoV-OC43感染,Wnt/β-catenin信号通路显着富集了BHK21细胞系的新合成转录本。此外,Wnt通路的抑制促进了病毒在感染早期的复制,但是在感染的后期抑制了它。此外,remesivir抑制HCoV-OC43早期感染诱导的Wnt/β-catenin信号通路的上调。总的来说,我们的研究表明Wnt/β-catenin通路在HCoV-OC43感染的不同阶段的不同作用,提示抗病毒治疗的潜在目标。此外,尽管HCoV-OC43感染在BHK21细胞中诱导细胞病变效应,抑制细胞凋亡不影响病毒的细胞内复制。基于这种时间分辨方法监测新合成的RNA是研究病毒感染机制的非常有前途的方法。
The progress of viral infection involves numerous transcriptional regulatory events. The identification of the newly synthesized transcripts helps us to understand the replication mechanisms and pathogenesis of the virus. Here, we utilized a time-resolved technique called metabolic RNA labeling approach called thiol(SH)-linked alkylation for the metabolic sequencing of RNA (SLAM-seq) to differentially elucidate the levels of steady-state and newly synthesized RNAs of BHK21 cell line in response to human coronavirus OC43 (HCoV-OC43) infection. Our results showed that the Wnt/β-catenin signaling pathway was significantly enriched with the newly synthesized transcripts of BHK21 cell line in response to HCoV-OC43 infection. Moreover, inhibition of the Wnt pathway promoted viral replication in the early stage of infection, but inhibited it in the later stage of infection. Furthermore, remdesivir inhibits the upregulation of the Wnt/β-catenin signaling pathway induced by early infection with HCoV-OC43. Collectively, our study showed the diverse roles of Wnt/β-catenin pathway at different stages of HCoV-OC43 infection, suggesting a potential target for the antiviral treatment. In addition, although infection with HCoV-OC43 induces cytopathic effects in BHK21 cells, inhibiting apoptosis does not affect the intracellular replication of the virus. Monitoring newly synthesized RNA based on such time-resolved approach is a highly promising method for studying the mechanism of viral infections.