Origins of Enterovirus Replication Organelles Established by Whole-Cell Electron Microscopy

ABSTRACT Enterovirus genome replication occurs at virus-induced structures derived from cellular membranes and lipids. However, the origin of these replication organelles (ROs) remains uncertain. Ultrastructural evidence of the membrane donor is lacking, suggesting that the sites of its transition i...

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Autores principales:	Charlotte E. Melia, Christopher J. Peddie, Anja W. M. de Jong, Eric J. Snijder, Lucy M. Collinson, Abraham J. Koster, Hilde M. van der Schaar, Frank J. M. van Kuppeveld, Montserrat Bárcena
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	replication organelle biogenesis coxsackievirus picornavirus serial block-face scanning electron microscopy SBF-SEM correlative light and electron microscopy Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/060c6b968b3649048287ffe1bae24e3f
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spelling	oai:doaj.org-article:060c6b968b3649048287ffe1bae24e3f2021-11-15T15:55:24ZOrigins of Enterovirus Replication Organelles Established by Whole-Cell Electron Microscopy10.1128/mBio.00951-192150-7511https://doaj.org/article/060c6b968b3649048287ffe1bae24e3f2019-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00951-19https://doaj.org/toc/2150-7511ABSTRACT Enterovirus genome replication occurs at virus-induced structures derived from cellular membranes and lipids. However, the origin of these replication organelles (ROs) remains uncertain. Ultrastructural evidence of the membrane donor is lacking, suggesting that the sites of its transition into ROs are rare or fleeting. To overcome this challenge, we combined live-cell imaging and serial block-face scanning electron microscopy of whole cells to capture emerging enterovirus ROs. The first foci of fluorescently labeled viral protein correlated with ROs connected to the endoplasmic reticulum (ER) and preceded the appearance of ROs stemming from the trans-Golgi network. Whole-cell data sets further revealed striking contact regions between ROs and lipid droplets that may represent a route for lipid shuttling to facilitate RO proliferation and genome replication. Our data provide direct evidence that enteroviruses use ER and then Golgi membranes to initiate RO formation, demonstrating the remarkable flexibility with which enteroviruses usurp cellular organelles. IMPORTANCE Enteroviruses are causative agents of a range of human diseases. The replication of these viruses within cells relies on specialized membranous structures termed replication organelles (ROs) that form during infection but whose origin remains elusive. To capture the emergence of enterovirus ROs, we use correlative light and serial block-face scanning electron microscopy, a powerful method to pinpoint rare events in their whole-cell ultrastructural context. RO biogenesis was found to occur first at ER and then at Golgi membranes. Extensive contacts were found between early ROs and lipid droplets (LDs), which likely serve to provide LD-derived lipids required for replication. Together, these data establish the dual origin of enterovirus ROs and the chronology of their biogenesis at different supporting cellular membranes.Charlotte E. MeliaChristopher J. PeddieAnja W. M. de JongEric J. SnijderLucy M. CollinsonAbraham J. KosterHilde M. van der SchaarFrank J. M. van KuppeveldMontserrat BárcenaAmerican Society for Microbiologyarticlereplication organelle biogenesiscoxsackieviruspicornavirusserial block-face scanning electron microscopySBF-SEMcorrelative light and electron microscopyMicrobiologyQR1-502ENmBio, Vol 10, Iss 3 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	replication organelle biogenesis coxsackievirus picornavirus serial block-face scanning electron microscopy SBF-SEM correlative light and electron microscopy Microbiology QR1-502
spellingShingle	replication organelle biogenesis coxsackievirus picornavirus serial block-face scanning electron microscopy SBF-SEM correlative light and electron microscopy Microbiology QR1-502 Charlotte E. Melia Christopher J. Peddie Anja W. M. de Jong Eric J. Snijder Lucy M. Collinson Abraham J. Koster Hilde M. van der Schaar Frank J. M. van Kuppeveld Montserrat Bárcena Origins of Enterovirus Replication Organelles Established by Whole-Cell Electron Microscopy
description	ABSTRACT Enterovirus genome replication occurs at virus-induced structures derived from cellular membranes and lipids. However, the origin of these replication organelles (ROs) remains uncertain. Ultrastructural evidence of the membrane donor is lacking, suggesting that the sites of its transition into ROs are rare or fleeting. To overcome this challenge, we combined live-cell imaging and serial block-face scanning electron microscopy of whole cells to capture emerging enterovirus ROs. The first foci of fluorescently labeled viral protein correlated with ROs connected to the endoplasmic reticulum (ER) and preceded the appearance of ROs stemming from the trans-Golgi network. Whole-cell data sets further revealed striking contact regions between ROs and lipid droplets that may represent a route for lipid shuttling to facilitate RO proliferation and genome replication. Our data provide direct evidence that enteroviruses use ER and then Golgi membranes to initiate RO formation, demonstrating the remarkable flexibility with which enteroviruses usurp cellular organelles. IMPORTANCE Enteroviruses are causative agents of a range of human diseases. The replication of these viruses within cells relies on specialized membranous structures termed replication organelles (ROs) that form during infection but whose origin remains elusive. To capture the emergence of enterovirus ROs, we use correlative light and serial block-face scanning electron microscopy, a powerful method to pinpoint rare events in their whole-cell ultrastructural context. RO biogenesis was found to occur first at ER and then at Golgi membranes. Extensive contacts were found between early ROs and lipid droplets (LDs), which likely serve to provide LD-derived lipids required for replication. Together, these data establish the dual origin of enterovirus ROs and the chronology of their biogenesis at different supporting cellular membranes.
format	article
author	Charlotte E. Melia Christopher J. Peddie Anja W. M. de Jong Eric J. Snijder Lucy M. Collinson Abraham J. Koster Hilde M. van der Schaar Frank J. M. van Kuppeveld Montserrat Bárcena
author_facet	Charlotte E. Melia Christopher J. Peddie Anja W. M. de Jong Eric J. Snijder Lucy M. Collinson Abraham J. Koster Hilde M. van der Schaar Frank J. M. van Kuppeveld Montserrat Bárcena
author_sort	Charlotte E. Melia
title	Origins of Enterovirus Replication Organelles Established by Whole-Cell Electron Microscopy
title_short	Origins of Enterovirus Replication Organelles Established by Whole-Cell Electron Microscopy
title_full	Origins of Enterovirus Replication Organelles Established by Whole-Cell Electron Microscopy
title_fullStr	Origins of Enterovirus Replication Organelles Established by Whole-Cell Electron Microscopy
title_full_unstemmed	Origins of Enterovirus Replication Organelles Established by Whole-Cell Electron Microscopy
title_sort	origins of enterovirus replication organelles established by whole-cell electron microscopy
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/060c6b968b3649048287ffe1bae24e3f
work_keys_str_mv	AT charlotteemelia originsofenterovirusreplicationorganellesestablishedbywholecellelectronmicroscopy AT christopherjpeddie originsofenterovirusreplicationorganellesestablishedbywholecellelectronmicroscopy AT anjawmdejong originsofenterovirusreplicationorganellesestablishedbywholecellelectronmicroscopy AT ericjsnijder originsofenterovirusreplicationorganellesestablishedbywholecellelectronmicroscopy AT lucymcollinson originsofenterovirusreplicationorganellesestablishedbywholecellelectronmicroscopy AT abrahamjkoster originsofenterovirusreplicationorganellesestablishedbywholecellelectronmicroscopy AT hildemvanderschaar originsofenterovirusreplicationorganellesestablishedbywholecellelectronmicroscopy AT frankjmvankuppeveld originsofenterovirusreplicationorganellesestablishedbywholecellelectronmicroscopy AT montserratbarcena originsofenterovirusreplicationorganellesestablishedbywholecellelectronmicroscopy
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Origins of Enterovirus Replication Organelles Established by Whole-Cell Electron Microscopy

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