Coronavirus RNA Synthesis Takes Place within Membrane-Bound Sites
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Viruses
2.2. Labeling of Nascent Viral RNA with Bromouridine
2.3. Immunofluorescence Labeling
3. Results
3.1. DsRNA Is Found in Membrane-Bound Compartments
3.2. DsRNA Is Closely Associated with Sites of Viral RNA Synthesis
3.3. Sites of Viral RNA Synthesis Are Membrane-Protected
3.4. Viral RNA Is Transported to the Cytoplasm Later in Infection
3.5. RNA Synthesis by All Genera of Coronaviruses Takes Place in a Membrane-Bound Compartment
4. Discussion
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Antibody Name in Text | Antibody Target | Source |
---|---|---|
dsRNA | J2 dsRNA | Scicons (English and Scientific Consulting, Budapest, Hungary) |
Nsp12 | IBV RdRp (nsp12) | Maier et al. mBio ‘13 |
IBV | IBV structural proteins (S, M, and N) | Abcam |
PDI | PDI | Enzo |
b-tubulin | b-tubulin | ThermoFisher |
BrU | Anti-Bromodeoxyuridine | Roche, Sigma |
229E N | HCoV 229E N protein | MRC PPU Reagents and Services |
SARS2 N | SARS-CoV2 N protein | MRC PPU Reagents and Services |
PDCoV N | PDCoV NP antigen | 2B Scientific |
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Doyle, N.; Simpson, J.; Hawes, P.C.; Maier, H.J. Coronavirus RNA Synthesis Takes Place within Membrane-Bound Sites. Viruses 2021, 13, 2540. https://doi.org/10.3390/v13122540
Doyle N, Simpson J, Hawes PC, Maier HJ. Coronavirus RNA Synthesis Takes Place within Membrane-Bound Sites. Viruses. 2021; 13(12):2540. https://doi.org/10.3390/v13122540
Chicago/Turabian StyleDoyle, Nicole, Jennifer Simpson, Philippa C. Hawes, and Helena J. Maier. 2021. "Coronavirus RNA Synthesis Takes Place within Membrane-Bound Sites" Viruses 13, no. 12: 2540. https://doi.org/10.3390/v13122540