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Three-Dimensional Remodeling of SARS-CoV2-Infected Cells Revealed by Cryogenic Soft X-ray Tomography
[Image: see text] Plus-strand RNA viruses are proficient at remodeling host cell membranes for optimal viral genome replication and the production of infectious progeny. These ultrastructural alterations result in the formation of viral membranous organelles and may be observed by different imaging...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10690842/ https://www.ncbi.nlm.nih.gov/pubmed/37939169 http://dx.doi.org/10.1021/acsnano.3c07265 |
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author | Castro, Victoria Pérez-Berna, Ana Joaquina Calvo, Gema Pereiro, Eva Gastaminza, Pablo |
author_facet | Castro, Victoria Pérez-Berna, Ana Joaquina Calvo, Gema Pereiro, Eva Gastaminza, Pablo |
author_sort | Castro, Victoria |
collection | PubMed |
description | [Image: see text] Plus-strand RNA viruses are proficient at remodeling host cell membranes for optimal viral genome replication and the production of infectious progeny. These ultrastructural alterations result in the formation of viral membranous organelles and may be observed by different imaging techniques, providing nanometric resolution. Guided by confocal and electron microscopy, this study describes the generation of wide-field volumes using cryogenic soft-X-ray tomography (cryo-SXT) on SARS-CoV-2-infected human lung adenocarcinoma cells. Confocal microscopy showed accumulation of double-stranded RNA (dsRNA) and nucleocapsid (N) protein in compact perinuclear structures, preferentially found around centrosomes at late stages of the infection. Transmission electron microscopy (TEM) showed accumulation of membranous structures in the vicinity of the infected cell nucleus, forming a viral replication organelle containing characteristic double-membrane vesicles and virus-like particles within larger vesicular structures. Cryo-SXT revealed viral replication organelles very similar to those observed by TEM but indicated that the vesicular organelle observed in TEM sections is indeed a vesiculo-tubular network that is enlarged and elongated at late stages of the infection. Overall, our data provide additional insight into the molecular architecture of the SARS-CoV-2 replication organelle. |
format | Online Article Text |
id | pubmed-10690842 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-106908422023-12-02 Three-Dimensional Remodeling of SARS-CoV2-Infected Cells Revealed by Cryogenic Soft X-ray Tomography Castro, Victoria Pérez-Berna, Ana Joaquina Calvo, Gema Pereiro, Eva Gastaminza, Pablo ACS Nano [Image: see text] Plus-strand RNA viruses are proficient at remodeling host cell membranes for optimal viral genome replication and the production of infectious progeny. These ultrastructural alterations result in the formation of viral membranous organelles and may be observed by different imaging techniques, providing nanometric resolution. Guided by confocal and electron microscopy, this study describes the generation of wide-field volumes using cryogenic soft-X-ray tomography (cryo-SXT) on SARS-CoV-2-infected human lung adenocarcinoma cells. Confocal microscopy showed accumulation of double-stranded RNA (dsRNA) and nucleocapsid (N) protein in compact perinuclear structures, preferentially found around centrosomes at late stages of the infection. Transmission electron microscopy (TEM) showed accumulation of membranous structures in the vicinity of the infected cell nucleus, forming a viral replication organelle containing characteristic double-membrane vesicles and virus-like particles within larger vesicular structures. Cryo-SXT revealed viral replication organelles very similar to those observed by TEM but indicated that the vesicular organelle observed in TEM sections is indeed a vesiculo-tubular network that is enlarged and elongated at late stages of the infection. Overall, our data provide additional insight into the molecular architecture of the SARS-CoV-2 replication organelle. American Chemical Society 2023-11-08 /pmc/articles/PMC10690842/ /pubmed/37939169 http://dx.doi.org/10.1021/acsnano.3c07265 Text en © 2023 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by-nc-nd/4.0/Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Castro, Victoria Pérez-Berna, Ana Joaquina Calvo, Gema Pereiro, Eva Gastaminza, Pablo Three-Dimensional Remodeling of SARS-CoV2-Infected Cells Revealed by Cryogenic Soft X-ray Tomography |
title | Three-Dimensional
Remodeling of SARS-CoV2-Infected
Cells Revealed by Cryogenic Soft X-ray Tomography |
title_full | Three-Dimensional
Remodeling of SARS-CoV2-Infected
Cells Revealed by Cryogenic Soft X-ray Tomography |
title_fullStr | Three-Dimensional
Remodeling of SARS-CoV2-Infected
Cells Revealed by Cryogenic Soft X-ray Tomography |
title_full_unstemmed | Three-Dimensional
Remodeling of SARS-CoV2-Infected
Cells Revealed by Cryogenic Soft X-ray Tomography |
title_short | Three-Dimensional
Remodeling of SARS-CoV2-Infected
Cells Revealed by Cryogenic Soft X-ray Tomography |
title_sort | three-dimensional
remodeling of sars-cov2-infected
cells revealed by cryogenic soft x-ray tomography |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10690842/ https://www.ncbi.nlm.nih.gov/pubmed/37939169 http://dx.doi.org/10.1021/acsnano.3c07265 |
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