Shedding light on reovirus assembly-Multimodal imaging of viral factories.

0583125 - BC 2024 RIV US eng J - Journal Article
Ďurinová, Eva - Mojzes, P. - Bílý, Tomáš - Franta, Z. - Fessl, T. - Borodavka, A. - Tůma, R.
Shedding light on reovirus assembly-Multimodal imaging of viral factories.
Advances in Virus Research. Roč. 116, JAN (2023), s. 173-213. ISSN 0065-3527. E-ISSN 1557-8399
R&D Projects: GA MŠMT(CZ) LM2023050; GA MŠMT(CZ) EF18_046/0016045
Institutional support: RVO:60077344
Keywords : RNA * Holography * Infection
OECD category: Cell biology
Impact factor: 9.938, year: 2021
Method of publishing: Limited access
https://www.sciencedirect.com/science/article/abs/pii/S0065352723000180?via%3Dihub

Avian (ortho)reovirus (ARV), which belongs to Reoviridae family, is a major domestic fowl pathogen and is the causative agent of viral tenosynovitis and chronic respiratory disease in chicken. ARV replicates within cytoplasmic inclusions, so-called viral factories, that form by phase separation and thus belong to a wider class of biological condensates. Here, we evaluate different optical imaging methods that have been developed or adapted to follow formation, fluidity and composition of viral factories and compare them with the complementary structural information obtained by well-established transmission electron microscopy and electron tomography. The molecular and cellular biology aspects for setting up and following virus infection in cells by imaging are described first. We then demonstrate that a wide-field version of fluorescence recovery after photobleaching is an effective tool to measure fluidity of mobile viral factories. A new technique, holotomographic phase microscopy, is then used for imaging of viral factory formation in live cells in three dimensions. Confocal Raman microscopy of infected cells provides chemical contrast for label-free segmentation of images and addresses important questions about biomolecular concentrations within viral factories and other biological condensates. Optical imaging is complemented by electron microscopy and tomography which supply higher resolution structural detail, including visualization of individual virions within the three-dimensional cellular context.
Permanent Link: https://hdl.handle.net/11104/0351117