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Ultrastructural insight into SARS-CoV-2 entry and budding in human airway epithelium

Ultrastructural studies of SARS-CoV-2 infected cells are crucial to better understand the mechanisms of viral entry and budding within host cells. Here, we examined human airway epithelium infected with three different isolates of SARS-CoV-2 including the B.1.1.7 variant by transmission electron mic...

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Autores principales: Pinto, Andreia L., Rai, Ranjit K., Brown, Jonathan C., Griffin, Paul, Edgar, James R., Shah, Anand, Singanayagam, Aran, Hogg, Claire, Barclay, Wendy S., Futter, Clare E., Burgoyne, Thomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8956608/
https://www.ncbi.nlm.nih.gov/pubmed/35338134
http://dx.doi.org/10.1038/s41467-022-29255-y
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author Pinto, Andreia L.
Rai, Ranjit K.
Brown, Jonathan C.
Griffin, Paul
Edgar, James R.
Shah, Anand
Singanayagam, Aran
Hogg, Claire
Barclay, Wendy S.
Futter, Clare E.
Burgoyne, Thomas
author_facet Pinto, Andreia L.
Rai, Ranjit K.
Brown, Jonathan C.
Griffin, Paul
Edgar, James R.
Shah, Anand
Singanayagam, Aran
Hogg, Claire
Barclay, Wendy S.
Futter, Clare E.
Burgoyne, Thomas
author_sort Pinto, Andreia L.
collection PubMed
description Ultrastructural studies of SARS-CoV-2 infected cells are crucial to better understand the mechanisms of viral entry and budding within host cells. Here, we examined human airway epithelium infected with three different isolates of SARS-CoV-2 including the B.1.1.7 variant by transmission electron microscopy and tomography. For all isolates, the virus infected ciliated but not goblet epithelial cells. Key SARS-CoV-2 entry molecules, ACE2 and TMPRSS2, were found to be localised to the plasma membrane including microvilli but excluded from cilia. Consistently, extracellular virions were seen associated with microvilli and the apical plasma membrane but rarely with ciliary membranes. Profiles indicative of viral fusion where tomography showed that the viral membrane was continuous with the apical plasma membrane and the nucleocapsids diluted, compared with unfused virus, demonstrate that the plasma membrane is one site of entry where direct fusion releasing the nucleoprotein-encapsidated genome occurs. Intact intracellular virions were found within ciliated cells in compartments with a single membrane bearing S glycoprotein. Tomography showed concentration of nucleocapsids round the periphery of profiles strongly suggestive of viral budding into these compartments and this may explain how virions gain their S glycoprotein containing envelope.
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spelling pubmed-89566082022-04-20 Ultrastructural insight into SARS-CoV-2 entry and budding in human airway epithelium Pinto, Andreia L. Rai, Ranjit K. Brown, Jonathan C. Griffin, Paul Edgar, James R. Shah, Anand Singanayagam, Aran Hogg, Claire Barclay, Wendy S. Futter, Clare E. Burgoyne, Thomas Nat Commun Article Ultrastructural studies of SARS-CoV-2 infected cells are crucial to better understand the mechanisms of viral entry and budding within host cells. Here, we examined human airway epithelium infected with three different isolates of SARS-CoV-2 including the B.1.1.7 variant by transmission electron microscopy and tomography. For all isolates, the virus infected ciliated but not goblet epithelial cells. Key SARS-CoV-2 entry molecules, ACE2 and TMPRSS2, were found to be localised to the plasma membrane including microvilli but excluded from cilia. Consistently, extracellular virions were seen associated with microvilli and the apical plasma membrane but rarely with ciliary membranes. Profiles indicative of viral fusion where tomography showed that the viral membrane was continuous with the apical plasma membrane and the nucleocapsids diluted, compared with unfused virus, demonstrate that the plasma membrane is one site of entry where direct fusion releasing the nucleoprotein-encapsidated genome occurs. Intact intracellular virions were found within ciliated cells in compartments with a single membrane bearing S glycoprotein. Tomography showed concentration of nucleocapsids round the periphery of profiles strongly suggestive of viral budding into these compartments and this may explain how virions gain their S glycoprotein containing envelope. Nature Publishing Group UK 2022-03-25 /pmc/articles/PMC8956608/ /pubmed/35338134 http://dx.doi.org/10.1038/s41467-022-29255-y Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Pinto, Andreia L.
Rai, Ranjit K.
Brown, Jonathan C.
Griffin, Paul
Edgar, James R.
Shah, Anand
Singanayagam, Aran
Hogg, Claire
Barclay, Wendy S.
Futter, Clare E.
Burgoyne, Thomas
Ultrastructural insight into SARS-CoV-2 entry and budding in human airway epithelium
title Ultrastructural insight into SARS-CoV-2 entry and budding in human airway epithelium
title_full Ultrastructural insight into SARS-CoV-2 entry and budding in human airway epithelium
title_fullStr Ultrastructural insight into SARS-CoV-2 entry and budding in human airway epithelium
title_full_unstemmed Ultrastructural insight into SARS-CoV-2 entry and budding in human airway epithelium
title_short Ultrastructural insight into SARS-CoV-2 entry and budding in human airway epithelium
title_sort ultrastructural insight into sars-cov-2 entry and budding in human airway epithelium
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8956608/
https://www.ncbi.nlm.nih.gov/pubmed/35338134
http://dx.doi.org/10.1038/s41467-022-29255-y
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