A spatial multi-scale fluorescence microscopy toolbox discloses entry checkpoints of SARS-CoV-2 variants in Vero E6 cells

We exploited a multi-scale microscopy imaging toolbox to address some major issues related to SARS-CoV-2 interactions with host cells. Our approach harnesses both conventional and super-resolution fluorescence microscopy and easily matches the spatial scale of single-virus/cell checkpoints. After it...

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Autores principales:	Barbara Storti, Paola Quaranta, Cristina Di Primio, Nicola Clementi, Nicasio Mancini, Elena Criscuolo, Pietro Giorgio Spezia, Vittoria Carnicelli, Giulia Lottini, Emanuele Paolini, Giulia Freer, Michele Lai, Mario Costa, Fabio Beltram, Alberto Diaspro, Mauro Pistello, Riccardo Zucchi, Paolo Bianchini, Giovanni Signore, Ranieri Bizzarri
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	SARS-CoV-2 spike B.1.1.7 variant of concern Late entry Clathrin STED dSTORM Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/38a5d9bc1ecb4185a14016ed78626b1d
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spelling	oai:doaj.org-article:38a5d9bc1ecb4185a14016ed78626b1d2021-11-26T04:26:43ZA spatial multi-scale fluorescence microscopy toolbox discloses entry checkpoints of SARS-CoV-2 variants in Vero E6 cells2001-037010.1016/j.csbj.2021.10.038https://doaj.org/article/38a5d9bc1ecb4185a14016ed78626b1d2021-01-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2001037021004608https://doaj.org/toc/2001-0370We exploited a multi-scale microscopy imaging toolbox to address some major issues related to SARS-CoV-2 interactions with host cells. Our approach harnesses both conventional and super-resolution fluorescence microscopy and easily matches the spatial scale of single-virus/cell checkpoints. After its validation through the characterization of infected cells and virus morphology, we leveraged this toolbox to reveal subtle issues related to the entry phase of SARS-CoV-2 variants in Vero E6 cells. Our results show that in Vero E6 cells the B.1.1.7 strain (aka Alpha Variant of Concern) is associated with much faster kinetics of endocytic uptake compared to its ancestor B.1.177. Given the cell-entry scenario dominated by the endosomal “late pathway”, the faster internalization of B.1.1.7 could be directly related to the N501Y mutation in the S protein, which is known to strengthen the binding of Spike receptor binding domain with ACE2. Remarkably, we also directly observed the central role of clathrin as a mediator of endocytosis in the late pathway of entry. In keeping with the clathrin-mediated endocytosis, we highlighted the non-raft membrane localization of ACE2. Overall, we believe that our fluorescence microscopy-based approach represents a fertile strategy to investigate the molecular features of SARS-CoV-2 interactions with cells.Barbara StortiPaola QuarantaCristina Di PrimioNicola ClementiNicasio ManciniElena CriscuoloPietro Giorgio SpeziaVittoria CarnicelliGiulia LottiniEmanuele PaoliniGiulia FreerMichele LaiMario CostaFabio BeltramAlberto DiasproMauro PistelloRiccardo ZucchiPaolo BianchiniGiovanni SignoreRanieri BizzarriElsevierarticleSARS-CoV-2 spikeB.1.1.7 variant of concernLate entryClathrinSTEDdSTORMBiotechnologyTP248.13-248.65ENComputational and Structural Biotechnology Journal, Vol 19, Iss , Pp 6140-6156 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	SARS-CoV-2 spike B.1.1.7 variant of concern Late entry Clathrin STED dSTORM Biotechnology TP248.13-248.65
spellingShingle	SARS-CoV-2 spike B.1.1.7 variant of concern Late entry Clathrin STED dSTORM Biotechnology TP248.13-248.65 Barbara Storti Paola Quaranta Cristina Di Primio Nicola Clementi Nicasio Mancini Elena Criscuolo Pietro Giorgio Spezia Vittoria Carnicelli Giulia Lottini Emanuele Paolini Giulia Freer Michele Lai Mario Costa Fabio Beltram Alberto Diaspro Mauro Pistello Riccardo Zucchi Paolo Bianchini Giovanni Signore Ranieri Bizzarri A spatial multi-scale fluorescence microscopy toolbox discloses entry checkpoints of SARS-CoV-2 variants in Vero E6 cells
description	We exploited a multi-scale microscopy imaging toolbox to address some major issues related to SARS-CoV-2 interactions with host cells. Our approach harnesses both conventional and super-resolution fluorescence microscopy and easily matches the spatial scale of single-virus/cell checkpoints. After its validation through the characterization of infected cells and virus morphology, we leveraged this toolbox to reveal subtle issues related to the entry phase of SARS-CoV-2 variants in Vero E6 cells. Our results show that in Vero E6 cells the B.1.1.7 strain (aka Alpha Variant of Concern) is associated with much faster kinetics of endocytic uptake compared to its ancestor B.1.177. Given the cell-entry scenario dominated by the endosomal “late pathway”, the faster internalization of B.1.1.7 could be directly related to the N501Y mutation in the S protein, which is known to strengthen the binding of Spike receptor binding domain with ACE2. Remarkably, we also directly observed the central role of clathrin as a mediator of endocytosis in the late pathway of entry. In keeping with the clathrin-mediated endocytosis, we highlighted the non-raft membrane localization of ACE2. Overall, we believe that our fluorescence microscopy-based approach represents a fertile strategy to investigate the molecular features of SARS-CoV-2 interactions with cells.
format	article
author	Barbara Storti Paola Quaranta Cristina Di Primio Nicola Clementi Nicasio Mancini Elena Criscuolo Pietro Giorgio Spezia Vittoria Carnicelli Giulia Lottini Emanuele Paolini Giulia Freer Michele Lai Mario Costa Fabio Beltram Alberto Diaspro Mauro Pistello Riccardo Zucchi Paolo Bianchini Giovanni Signore Ranieri Bizzarri
author_facet	Barbara Storti Paola Quaranta Cristina Di Primio Nicola Clementi Nicasio Mancini Elena Criscuolo Pietro Giorgio Spezia Vittoria Carnicelli Giulia Lottini Emanuele Paolini Giulia Freer Michele Lai Mario Costa Fabio Beltram Alberto Diaspro Mauro Pistello Riccardo Zucchi Paolo Bianchini Giovanni Signore Ranieri Bizzarri
author_sort	Barbara Storti
title	A spatial multi-scale fluorescence microscopy toolbox discloses entry checkpoints of SARS-CoV-2 variants in Vero E6 cells
title_short	A spatial multi-scale fluorescence microscopy toolbox discloses entry checkpoints of SARS-CoV-2 variants in Vero E6 cells
title_full	A spatial multi-scale fluorescence microscopy toolbox discloses entry checkpoints of SARS-CoV-2 variants in Vero E6 cells
title_fullStr	A spatial multi-scale fluorescence microscopy toolbox discloses entry checkpoints of SARS-CoV-2 variants in Vero E6 cells
title_full_unstemmed	A spatial multi-scale fluorescence microscopy toolbox discloses entry checkpoints of SARS-CoV-2 variants in Vero E6 cells
title_sort	spatial multi-scale fluorescence microscopy toolbox discloses entry checkpoints of sars-cov-2 variants in vero e6 cells
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/38a5d9bc1ecb4185a14016ed78626b1d
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A spatial multi-scale fluorescence microscopy toolbox discloses entry checkpoints of SARS-CoV-2 variants in Vero E6 cells

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