Ferreira-Venter, LindaVenter, EudriTheron, JacquesVan Staden, Vida2019-07-242019-05Ferreira-Venter, L., Venter, E., Theron, J. et al. 2019, 'Targeted mutational analysis to unravel the complexity of African horse sickness virus NS3 function in mammalian cells', Virology, vol. 531, pp. 149-161.0042-6822 (print)1096-0341 (online)10.1016/j.virol.2019.03.005http://hdl.handle.net/2263/70784The African horse sickness virus non-structural protein 3 (NS3) is involved in the final stages of infection. To gain insight into the function of different NS3 domains, we generated reverse genetics-derived mutants, each expressing a modified version of the protein. A functional comparison of these mutants to the wild-type virus in mammalian cells indicated the variable contribution of the different domains to the cytopathic effect and in ensuring effective virus trafficking and release. The transmembrane domains were determined as essential mediators of NS3 localisation, as the abnormal processing of these mutant proteins resulted in their nuclear localisation and interaction with NS1. NS3 cytoplasmic domain disruptions resulted in increased cytosolic virus particle accumulation and abnormal virion tethering to plasma membranes. Other aspects of infection were also affected, such as VIB formation and distribution of the outer capsid proteins. Overall, these results illustrate the intricate role of NS3 in the infection cycle.en© 2019 Elsevier Inc. All rights reserved. Notice : this is the author’s version of a work that was accepted for publication in Virology. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. A definitive version was subsequently published in Virology, vol. 531, pp. 149-161, 2019. doi : 10.1016/j.virol.2019.03.005.Rous sarcoma virus (RSV)PathwayReleaseGagTraffickingLipid raftsInsect cellAfrican horsesickness virus (AHSV)Bluetongue virus (BTV)Non-structural protein 3 (NS3)Reverse geneticsVirus traffickingVirus releaseNon-structural protein 1 (NS1)Protein functional domainsPostprint Article