Insight into the replication of RNA viruses by spatiotemporal analyses of virus-host interaction

Emerging and re-emerging viruses, especially the RNA viruses, are rising treats to human as well as other species in recent years. Despite the rapid advances in techniques used to identify and detect emerging viruses, comprehensively analyzing and understanding the mechanism of virus replication still has the pivotal role in antiviral development. Because viruses can only complete their life cycle within the host cells, characterizing the viral-host interactions will provide the fundamental information of viral replication. Here, we design an assay platform to systematically investigate the replication of positive-stranded RNA virus based on the metabolic labeling. Since the material to be labeled is the replicating genome, all RNA viruses capable of infecting the cells are supposed to be studied using methods described in this proposal. Full-genome labeling will be achieved through 4-thioluridine incorporation during viral replication, and viral RNA-associated protein complexes (vRNP) at different stages of replication can be assessed. In addition to revealing the vRNP remodeling during viral replication, we will also examine the host responses in an infection-specific manner. Newly synthesized RNAs, both coding and non-coding RNA, stimulated by infection will be collected and sequenced, and their importance on viral replication will be explored. Finally, the features of the temporal RNPs/RNAs identified through metabolic labeling at the cellular level will be studied by subcellular fractionation and in situ hybridization experiments. Thus the basic principle in respect of the spatiotemporal regulation of an emerging virus infection can be elucidated through the pipeline.

Project ID:PC10910-0006
External Project ID:MOST109-2320-B182-045-MY2