Expression of the Heterotrimeric GP2/GP3/GP4 Spike of an Arterivirus in Mammalian Cells

Expression of the Heterotrimeric GP2/GP3/GP4 Spike of an Arterivirus in Mammalian Cells

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Equine arteritis virus (EAV), an enveloped positive-strand RNA virus, is an important pathogen of horses and the prototype member of the Arteiviridae family.Unlike many other enveloped viruses, which possess homotrimeric spikes, the spike responsible Human iPSC-derived astrocytes generated from donors with globoid cell leukodystrophy display phenotypes associated with disease for cellular tropism in Arteriviruses is a heterotrimer composed of 3 glycoproteins: GP2, GP3, and GP4.Together with the hydrophobic protein E they are the minor components of virus particles.We describe the expression of all 3 minor glycoproteins, each equipped with a different tag, from a multi-cassette system in mammalian BHK-21 cells.

Coprecipitation studies suggest that a rather small faction of GP2, GP3, and GP4 form dimeric or trimeric complexes.GP2, GP3, and GP4 co-localize with each other and also, albeit weaker, with the E-protein.The co-localization of GP3-HA and GP2-myc was tested with markers for ER, ERGIC, and cis-Golgi.The co-localization of GP3-HA was the same regardless of whether it was expressed alone or as a complex, whereas the transport of GP2-myc to cis-Golgi was higher when this protein was expressed as a complex.

The glycosylation pattern was also independent of whether the proteins were expressed alone or together.The recombinant spike might be a tool for basic research but might also be “Você é o que Google diz que você é”: a vida editável, entre controle e espetáculo used as a subunit vaccine for horses.