Abstract
Vaccinia virus relies on a series of proteolytic cleavage events involving two viral proteins, I7 and G1, to complete its life cycle. Furthermore, G1 itself is cleaved during vaccinia virus infection. However, convincing evidence is lacking to show whether G1 participates in autoproteolysis or is a substrate of another protease. We employed both biochemical and cell-based approaches to investigate G1 cleavage. G1, when expressed in bacteria, rabbit reticulocyte lysates or HeLa cells, was not processed. Moreover, G1 was cleaved in infected cells, but only in the presence of virus late gene expression; cleavage was strongly inhibited by proteasome inhibitors. Thus, these results imply a more complex G1 cleavage reaction than previously envisaged.
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Acknowledgements
We thank Dr. Bernard Moss from the National Institutes of Health (NIH), USA, for kindly providing us with a plasmid containing the G1 ORF and with the vaccinia virus conditional lethal mutant for G1L (vG1Li), Irene Gösler for support with cell culture, and the Bergthaler group at CeMM, Vienna, for support with the VACV experiments.
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This study was supported by the Austrian Science Fund (FWF) grant number M-1342-B13 to F.G.G.L.
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Leite, F.G.G., Bergthaler, A. & Skern, T. Vaccinia virus G1 protein: absence of autocatalytic self-processing. Arch Virol 162, 2803–2808 (2017). https://doi.org/10.1007/s00705-017-3409-y
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DOI: https://doi.org/10.1007/s00705-017-3409-y