In-situ cryo-electron tomography of Enterovirus replication
| Authors | |
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| Year of publication | 2023 |
| Type | Conference abstract |
| MU Faculty or unit | |
| Citation | |
| Description | Enteroviruses, a large group of non-enveloped picornaviruses, cause a range of human diseases, from the common cold to poliomyelitis. The molecular mechanisms of enterovirus replication and virion assembly are not well understood. Here, we present the cryo-FIB milling and cryo-electron tomography of Echovirus 18-infected cells to characterize the late stages of enterovirus infection in situ. We describe rearrangements of cellular membranes that enable the formation of viral replication factories. Furthermore, we have identified different virus particle assembly intermediates – capsid segments, open and complete capsids, and progeny virions. Some of the virus particles and assembly intermediates were localized in the cytoplasm, whereas others were bound to membranes. We have employed template matching using the structures of the whole virus capsid and a capsid missing three pentamers to identify the complete genome-containing and empty virus particles and virus assembly intermediates, respectively. The particles have been further analysed by subtomogram averaging and classification. Our results provide insight into the virus arrangement during the late stages of the replication cycle of enteroviruses. Additionally, the employed subtomogram averaging pipeline presents a further analysis of viruses and large molecular complexes in situ. We acknowledge the Cryo-electron microscopy and Tomography Core Facility of CEITEC MU for their support in data collection and analysis. |
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