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Viruses2022; 14(4); doi: 10.3390/v14040749

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

Abstract: 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 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 used as a subunit vaccine for horses.
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Publication Date: 2022-04-01 PubMed ID: 35458479PubMed Central: PMC9030998DOI: 10.3390/v14040749Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research explores the expression of three minor glycoproteins (GP2, GP3, and GP4), which form a heterotrimeric spike responsible for cellular tropism in arteriviruses, in BHK-21 mammalian cells. The findings indicate that the proteins co-localize with each other and the E-protein, and their glycosylation pattern remains unaffected whether they are expressed alone or together. This research could potentially be crucial in creating subunit vaccines for horses.

Arterivirus and Its Key Components

  • The study focuses on equine arteritis virus (EAV), a positive-strand RNA virus enveloped by a protective layer. EAV is widely recognized for causing diseases in horses and belongs to the Arteiviridae family.
  • Unlike other enveloped viruses that have homotrimeric spikes, arteriviruses possess heterotrimeric spikes composed of glycoproteins GP2, GP3, and GP4.
  • The glycoproteins, along with the hydrophobic E-protein, are minor but key components of the virus particles.

Expression of Glycoproteins in Mammalian Cells

  • The research describes the expression of the three glycoproteins, each with a unique tag, from a multi-cassette system in mammalian BHK-21 cells.
  • The study involves coprecipitation explorations, hinting at the possibility of the three glycoproteins forming dimeric or trimeric complexes.
  • The results reveal that the glycoproteins co-localize not only amongst each other but, albeit weakly, with the E-protein as well.

Co-localization and Transport of Glycoproteins

  • Markers for ER, ERGIC, and cis-Golgi were used to test the co-localization of GP3-HA and GP2-myc.
  • The co-localization of GP3-HA remained the same, irrespective of whether it was expressed individually or as a complex.
  • It was observed that the transport of GP2-myc to cis-Golgi was higher when expressed as a complex compared to when expressed alone.

Glycosylation Pattern and Practical Application

  • The glycosylation pattern exhibited by the proteins was unchanged, regardless of whether the glycoproteins were expressed individually or in combination.
  • This research suggests that the recombinant spike, derived from these glycoproteins, could be significantly beneficial for further basic research.
  • More importantly, due to the vaccines’ specificity to horses and the role of these glycoproteins in arteriviruses, this research could impact the development of subunit vaccines for horses in the future.

Cite This Article

APA
Matczuk AK, Zhang M, Veit M, Ugorski M. (2022). Expression of the Heterotrimeric GP2/GP3/GP4 Spike of an Arterivirus in Mammalian Cells. Viruses, 14(4). https://doi.org/10.3390/v14040749

Publication

Viruses
ISSN: 1999-4915
NlmUniqueID: 101509722
Country: Switzerland
Language: English
Volume: 14
Issue: 4

Researcher Affiliations

Matczuk, Anna Karolina
  • Division of Microbiology, Department of Pathology, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, 50-375 Wrocław, Poland.
Zhang, Minze
  • Institute of Virology, Department of Veterinary Medicine, Free University Berlin, 14163 Berlin, Germany.
Veit, Michael
  • Institute of Virology, Department of Veterinary Medicine, Free University Berlin, 14163 Berlin, Germany.
Ugorski, Maciej
  • Department of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, 50-375 Wrocław, Poland.

MeSH Terms

  • Animals
  • Arterivirus
  • Equartevirus / genetics
  • Equartevirus / metabolism
  • Glycoproteins / genetics
  • Guanidines
  • Horses
  • Mammals
  • Piperazines
  • Viral Envelope Proteins / metabolism

Conflict of Interest Statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

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