Translation and Replication Dynamics of Single RNA Viruses

RNA viruses are among the most prevalent pathogens and are a major burden on society. Although RNA viruses have been studied extensively, little is known about the processes that occur during the first several hours of infection because of a lack of sensitive assays. Here we develop a single-molecul...

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Bibliographic Details
Published in:Cell 2020-12, Vol.183 (7), p.1930-1945.e23
Main Authors: Boersma, Sanne, Rabouw, Huib H., Bruurs, Lucas J.M., Pavlovič, Tonja, van Vliet, Arno L.W., Beumer, Joep, Clevers, Hans, van Kuppeveld, Frank J.M., Tanenbaum, Marvin E.
Format: Article
Language:English
Subjects:
Cell Line, Tumor
Cell Survival
HEK293 Cells
Host-Pathogen Interactions
Humans
Interferons - metabolism
Protein Biosynthesis
Reproducibility of Results
Resource
RNA Transport
RNA virus
RNA Viruses - physiology
RNA, Viral - genetics
Single Molecule Imaging
SunTag
Time Factors
viral replication
viral translation
VIRIM
Virus Replication - physiology
virus-host competition
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Summary:RNA viruses are among the most prevalent pathogens and are a major burden on society. Although RNA viruses have been studied extensively, little is known about the processes that occur during the first several hours of infection because of a lack of sensitive assays. Here we develop a single-molecule imaging assay, virus infection real-time imaging (VIRIM), to study translation and replication of individual RNA viruses in live cells. VIRIM uncovered a striking heterogeneity in replication dynamics between cells and revealed extensive coordination between translation and replication of single viral RNAs. Furthermore, using VIRIM, we identify the replication step of the incoming viral RNA as a major bottleneck of successful infection and identify host genes that are responsible for inhibition of early virus replication. Single-molecule imaging of virus infection is a powerful tool to study virus replication and virus-host interactions that may be broadly applicable to RNA viruses. [Display omitted] •Single-molecule imaging assay to study translation and replication of +RNA viruses•Early picornavirus infection occurs in five distinct phases•Heterogeneity in translation and replication of viral RNAs•Replication of incoming vRNA represents major target for host antiviral activity Boersma et al. develop a single-molecule imaging assay (VIRIM) to study translation, replication, and virus-host interactions of +RNA viruses. They observe heterogeneity in translation and replication of single viruses, identify replication of the incoming viral genome as a bottleneck for successful infection, and identify host genes mediating this antiviral activity.
ISSN:0092-8674
1097-4172
DOI:10.1016/j.cell.2020.10.019