Imperial College London
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Professor Michael A. ("Mike") Skinner

Faculty of Medicine, Department of Infectious Disease

Emeritus Professor in Virology
 
 
 
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Contact

 

+44 (0)20 7594 3938m.skinner Website

 
 
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Assistant

 

Mrs Yasmin Mallu +44 (0)20 7594 3972

 
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Location

 

315Medical SchoolSt Mary's Campus

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Summary

 

Publications

Citation

BibTex format

@article{Campbell:2020:10.1128/JVI.02107-19,
author = {Campbell, EA and Reddy, VRAP and Gray, AG and Wells, J and Simpson, J and Skinner, MA and Hawes, PC and Broadbent, AJ},
doi = {10.1128/JVI.02107-19},
journal = {Journal of Virology},
pages = {1--16},
title = {Discrete virus factories form in the cytoplasm of cells co-infected with two replication competent tagged reporter birnaviruses, that subsequently coalesce over time.},
url = {http://dx.doi.org/10.1128/JVI.02107-19},
volume = {94},
year = {2020}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - The Birnaviridae family, responsible for major economic losses to poultry and aquaculture, are non-enveloped viruses with a segmented double-stranded (ds)RNA genome that replicate in discrete cytoplasmic virus factories (VFs). Reassortment is common, however, the underlying mechanism remains unknown given that VFs may act as a barrier to genome mixing. In order to provide new information on VF trafficking during dsRNA virus co-infection, we rescued two recombinant infectious bursal disease viruses (IBDVs) of strain PBG98 containing either a split GFP11- or Tetracysteine (TC)- tag fused to the VP1 polymerase (PBG98-VP1-GFP11 and PBG98-VP1-TC). DF-1 cells transfected with GFP1-10 prior to PBG98-VP1-GFP11 infection, or stained with ReAsH following PBG98-VP1-TC infection, had green or red foci in the cytoplasm respectively that co-localised with VP3 and dsRNA, consistent with VFs. The average number of VFs decreased from a mean of 60 to 5 per cell between 10 and 24 hours post infection (hpi) (p<0.0001), while the average area increased from 1.24 μm2 to 45.01μm2 (p<0.0001), and live cell imaging revealed that the VFs were highly dynamic structures that coalesced in the cytoplasm. Small VFs moved faster than large (average 0.57μm/s at 16 hpi compared to 0.22 μm/s at 22 hpi), and VF coalescence was dependent on an intact microtubule network and actin cytoskeleton. During co-infection with PBG98-VP1-GFP11 and PBG98-VP1-TC viruses, discrete VFs initially formed from each input virus that subsequently coalesced 10-16 hpi, and we speculate that Birnaviridae reassortment requires VF coalescence.IMPORTANCE Reassortment is common in viruses with segmented double stranded (ds)RNA genomes. However, these viruses typically replicate within discrete cytoplasmic virus factories (VFs) that may represent a barrier to genome mixing. We generated the first replication competent tagged reporter birnaviruses, infectious bursal disease viruses (IBDVs) containing a split GFP
AU  - Campbell,EA
AU  - Reddy,VRAP
AU  - Gray,AG
AU  - Wells,J
AU  - Simpson,J
AU  - Skinner,MA
AU  - Hawes,PC
AU  - Broadbent,AJ
DO  - 10.1128/JVI.02107-19
EP  - 16
PY  - 2020///
SN  - 0022-538X
SP  - 1
TI  - Discrete virus factories form in the cytoplasm of cells co-infected with two replication competent tagged reporter birnaviruses, that subsequently coalesce over time.
T2  - Journal of Virology
UR  - http://dx.doi.org/10.1128/JVI.02107-19
UR  - https://www.ncbi.nlm.nih.gov/pubmed/32321810
UR  - https://jvi.asm.org/content/early/2020/04/16/JVI.02107-19
UR  - http://hdl.handle.net/10044/1/79079
VL  - 94
ER  -
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