Establishment of a reverse genetics system for Schmallenberg virus, a newly emerged orthobunyavirus in Europe

Elliott, R.M.; Blakqori, G.; van Knippenberg, I.C.; Koudriakova, E.; Li, P.; McLees, A.; Shi, X.; Szemiel, A.M.

Abstract

Schmallenberg virus (SBV) is a newly emerged orthobunyavirus that has caused widespread disease in cattle, sheep and goats in Europe. Like other orthobunyaviruses, SBV is characterized by a tripartite negative-sense RNA genome that encodes four structural and two non-structural proteins. This study showed that SBV has a wide in vitro host range, and that BHK-21 cells are a convenient host for both SBV propagation and assay by plaque titration. The SBV genome segments were cloned as cDNA and a three-plasmid rescue system was established to recover infectious virus. Recombinant virus behaved similarly in cell culture to authentic virus. The ORF for the non-structural NSs protein, encoded on the smallest genome segment, was disrupted by introduction of translation stop codons in the appropriate cDNA, and when this plasmid was used in reverse genetics, a recombinant virus that lacked NSs expression was recovered. This virus had reduced capacity to shut-off host-cell protein synthesis compared with the wild-type virus. In addition, the NSs-deleted virus induced interferon (IFN) in cells, indicating that, like other orthobunyaviruses, NSs functions as an IFN antagonist, most probably by globally inhibiting host-cell metabolism. The development of a robust reverse genetics system for SBV will facilitate investigation of its pathogenic mechanisms as well as the creation of attenuated strains that could be candidate vaccines.

Citation

Elliott, R., Blakqori, G., van Knippenberg, I., Koudriakova, E., Li, P., McLees, A., …Szemiel, A. (2013). Establishment of a reverse genetics system for Schmallenberg virus, a newly emerged orthobunyavirus in Europe. Journal of General Virology, 94(4), https://doi.org/10.1099/vir.0.049981-0

• Journal Article Type: Article
• Acceptance Date: Dec 18, 2012
• Publication Date: 2013-04
• Deposit Date: Jul 19, 2023
• Journal: The Journal of general virology
• Print ISSN: 0022-1317
• Publisher: Microbiology Society
• Peer Reviewed: Peer Reviewed
• Volume: 94
• Issue: 4
• DOI: https://doi.org/10.1099/vir.0.049981-0