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. 2022 Jan 22;10(2):235. doi: 10.3390/biomedicines10020235

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Replication of VACV expressing distinct mutants of E3 in JC cells. A multi-step growth curve was conducted for 72 h in JC cells infected at a multiplicity of infection (MOI) of 0.01. The output virus was normalized against the input virus and graphed. VACV expressing E3 mutants unable to bind to dsRNA (E3LΔ26C) and Z-DNA (E3LΔ83N, E3L Y48A, E3L P63A and partially impaired mutants E3L Y48F and E3L P64A) displayed limited ability to replicate in JC cells. Mutations in residues not associated with Z-DNA-binding (E42A and Y57A) did not appreciably affect replication in JC cells. This data demonstrates the importance of both the N- and C-terminal domains of E3 for virus replication in JC cells and it corresponds to what was seen in vivo (see the main text).