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Production of Recombinant EAV with Tagged Structural Protein Gp3 to Study Artervirus Minor Protein Localization in Infected Cells

1
Department of Pathology, Division of Microbiology, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Wrocław 50-375, Poland
2
Department of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Wrocław 50-375, Poland
3
Confocal Microscopy Laboratory, PORT Polish Center for Technology Development, Wrocław 54-066, Poland
*
Author to whom correspondence should be addressed.
Viruses 2019, 11(8), 735; https://doi.org/10.3390/v11080735
Received: 5 July 2019 / Revised: 29 July 2019 / Accepted: 7 August 2019 / Published: 9 August 2019
(This article belongs to the Section Animal Viruses)
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Abstract

Equine arteritis virus (EAV) is a prototype member of the Arterivirus family, comprising important pathogens of domestic animals. Minor glycoproteins of Arteriviruses are responsible for virus entry and cellular tropism. The experimental methods for studying minor Arterivirus proteins are limited because of the lack of antibodies and nested open reading frames (ORFs). In this study, we generated recombinant EAV with separated ORFs 3 and 4, and Gp3 carrying HA-tag (Gp3-HA). The recombinant viruses were stable on passaging and replicated in titers similar to the wild-type EAV. Gp3-HA was incorporated into the virion particles as monomers and as a Gp2/Gp3-HA/Gp4 trimer. Gp3-HA localized in ER and, to a lesser extent, in the Golgi, it also co-localized with the E protein but not with the N protein. The co-localization of Gp3-HA and the E protein with ERGIC was reduced. Moreover, EAV with Gp3-HA could become a valuable research tool for identifying host cell factors during infection and the role of Gp3 in virus attachment and entry. View Full-Text
Keywords: EAV; arterivirus; Gp3; tagged recombinant virus; co-localization; trafficking EAV; arterivirus; Gp3; tagged recombinant virus; co-localization; trafficking
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Matczuk, A.K.; Chodaczek, G.; Ugorski, M. Production of Recombinant EAV with Tagged Structural Protein Gp3 to Study Artervirus Minor Protein Localization in Infected Cells. Viruses 2019, 11, 735.

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