Translation and Replication Dynamics of Single RNA Viruses
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
(2020) Cell, volume 183, issue 7, pp. 1930 - 1945 e1-13
(Article)
Abstract
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
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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.
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Keywords: +RNA virus, SunTag, VIRIM, single-molecule imaging, viral replication, viral translation, virus-host competition, General Biochemistry,Genetics and Molecular Biology
ISSN: 0092-8674
Publisher: Cell Press
Note: Funding Information: We thank members of the Tanenbaum and Van Kuppeveld labs for helpful discussions. We also thank Ive Logister, Stijn Sonneveld, and Rupa Banerjee for help with experiments. This work was financially supported by an ERC starting grant to M.E.T. ( EU/ERC-677936 RNAREG ), the Howard Hughes Medical Institute through an international research scholar grant to M.E.T. ( HHMI/IRS 55008747 ), and a NWO VICI grant to F.J.M.v.K. ( 91812628 ). S.B., H.H.R., L.J.M.B., T.P., J.B., H.C., and M.E.T. were supported by the Oncode Institute, which is partly funded by the Dutch Cancer Society (KWF). Publisher Copyright: © 2020 The Author(s) Copyright: Copyright 2021 Elsevier B.V., All rights reserved.
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