Infectious bronchitis virus generates spherules from zippered endoplasmic reticulum membranes

Maier, Helena J, Hawes, Philippa C., Cottam, Eleanor M., Mantell, Judith, Verkade, Paul, Monaghan, Paul, Wileman, Tom and Britton, Paul (2013) Infectious bronchitis virus generates spherules from zippered endoplasmic reticulum membranes. mBIO, 4 (5). ISSN 2150-7511

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Abstract

Replication of positive-sense RNA viruses is associated with the rearrangement of cellular membranes. Previous work on the infection of tissue culture cell lines with the betacoronaviruses mouse hepatitis virus and severe acute respiratory syndrome coronavirus (SARS-CoV) showed that they generate double-membrane vesicles (DMVs) and convoluted membranes as part of a reticular membrane network. Here we describe a detailed study of the membrane rearrangements induced by the avian gammacoronavirus infectious bronchitis virus (IBV) in a mammalian cell line but also in primary avian cells and in epithelial cells of ex vivo tracheal organ cultures. In all cell types, structures novel to IBV infection were identified that we have termed zippered endoplasmic reticulum (ER) and spherules. Zippered ER lacked luminal space, suggesting zippering of ER cisternae, while spherules appeared as uniform invaginations of zippered ER. Electron tomography showed that IBV-induced spherules are tethered to the zippered ER and that there is a channel connecting the interior of the spherule with the cytoplasm, a feature thought to be necessary for sites of RNA synthesis but not seen previously for membrane rearrangements induced by coronaviruses. We also identified DMVs in IBV-infected cells that were observed as single individual DMVs or were connected to the ER via their outer membrane but not to the zippered ER. Interestingly, IBV-induced spherules strongly resemble confirmed sites of RNA synthesis for alphaviruses, nodaviruses, and bromoviruses, which may indicate similar strategies of IBV and these diverse viruses for the assembly of RNA replication complexes

Item Type:	Article
Additional Information:	Copyright © 2013 Maier et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.
Uncontrolled Keywords:	mouse hepatitis-virus,coronavirus rna-synthesis,tracheal organ-cultures,semliki-forest-virus,replication complex,ultrastructural characterization,electron-microscope,arbovirus infection,sars-coronavirus,cells,sdg 3 - good health and well-being ,/dk/atira/pure/sustainabledevelopmentgoals/good_health_and_well_being
Faculty \ School:	Faculty of Medicine and Health Sciences > Norwich Medical School
UEA Research Groups:	Faculty of Medicine and Health Sciences > Research Groups > Gastroenterology and Gut Biology Faculty of Medicine and Health Sciences > Research Centres > Lifespan Health
Depositing User:	Pure Connector
Date Deposited:	12 Nov 2014 16:46
Last Modified:	19 Oct 2023 01:23
URI:	https://ueaeprints.uea.ac.uk/id/eprint/50785
DOI:	10.1128/mBio.00801-13

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