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Virology2001; 288(2); 283-294; doi: 10.1006/viro.2001.1074

Construction of chimeric arteriviruses reveals that the ectodomain of the major glycoprotein is not the main determinant of equine arteritis virus tropism in cell culture.

Abstract: The recent development of arterivirus full-length cDNA clones makes possible the construction of chimeric arteriviruses for fundamental and applied studies. Using an equine arteritis virus (EAV) infectious cDNA clone, we have engineered chimeras in which the ectodomains of the two major envelope proteins, the glycoprotein GP(5) and the membrane protein M, were replaced by sequences from envelope proteins of related and unrelated RNA viruses. Using immunofluorescence microscopy, we monitored the transport of the hybrid GP(5) and M proteins to the Golgi complex, which depends on their heterodimerization and is a prerequisite for virus assembly. The only viable chimeras were those containing the GP(5) ectodomain from the porcine (PRRSV) or mouse (LDV) arteriviruses, which are both considerably smaller than the corresponding sequence of EAV. Although the two viable GP(5) chimeras were attenuated, they were still able to infect baby hamster kidney (BHK-21) and rabbit kidney (RK-13) cells. These cells can be infected by EAV, but not by either PRRSV or LDV. This implies that the ectodomain of the major glycoprotein GP(5), which has been postulated to be involved in receptor recognition, is not the main determinant of EAV tropism in cell culture.
Copyright 2001 Academic Press.
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Publication Date: 2001-10-17 PubMed ID: 11601900DOI: 10.1006/viro.2001.1074Google Scholar: Lookup
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Summary

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The research article describes the creation of chimeric arteriviruses to study the role of the major glycoprotein ectodomain in equine arteritis virus (EAV) cell culture tropism. The findings suggest that the ectodomain is not the primary determinant of EAV’s tropism.

Research Methodology

The researchers used a distinct method for their study:

  • The full-length cDNA clones of arteriviruses were utilized to create chimeric arteriviruses. These chimeras were created using an equine arteritis virus (EAV) infectious cDNA clone.
  • The ectodomains of the two major envelope proteins, glycoprotein GP(5) and the membrane protein M, were replaced with sequences derived from envelope proteins of related and unrelated RNA viruses.
  • Using immunofluorescence microscopy, the transport of the hybrid GP(5) and M proteins to the Golgi complex was monitored. Their heterodimerization influenced this transport, which is a crucial step for virus assembly.

Results and Findings

The researchers were able to obtain significant results:

  • The chimeras containing the GP(5) ectodomain from porcine (PRRSV) or mouse (LDV) arteriviruses were the only viable ones. The sequences from these arteriviruses were considerably smaller than the corresponding sequence of EAV.
  • Even though the successful GP(5) chimeras were weakened, they could still infect baby hamster kidney (BHK-21) and rabbit kidney (RK-13) cells. These cells are susceptible to EAV infection, but not to either PRRSV or LDV.

Conclusion

In conclusion, the major glycoprotein GP(5)’s ectodomain, previously hypothesized to involve receptor recognition is not the main determinant of EAV tropism in cell culture. This conclusion is based on the observation that chimeras could infect cell types usually not susceptible to their original viruses, implying other factors beyond the GP(5) ectodomain determine EAV tropism.

Cite This Article

APA
Dobbe JC, van der Meer Y, Spaan WJ, Snijder EJ. (2001). Construction of chimeric arteriviruses reveals that the ectodomain of the major glycoprotein is not the main determinant of equine arteritis virus tropism in cell culture. Virology, 288(2), 283-294. https://doi.org/10.1006/viro.2001.1074

Publication

Virology
ISSN: 0042-6822
NlmUniqueID: 0110674
Country: United States
Language: English
Volume: 288
Issue: 2
Pages: 283-294

Researcher Affiliations

Dobbe, J C
  • Department of Virology, Center of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.
van der Meer, Y
    Spaan, W J
      Snijder, E J

        MeSH Terms

        • Amino Acid Sequence
        • Animals
        • Binding Sites
        • Biological Transport
        • Cell Line
        • Cricetinae
        • Dimerization
        • Equartevirus / genetics
        • Equartevirus / metabolism
        • Horses
        • Lactate dehydrogenase-elevating virus / genetics
        • Mice
        • Molecular Sequence Data
        • Porcine respiratory and reproductive syndrome virus / genetics
        • Recombination, Genetic
        • Swine
        • Viral Envelope Proteins / genetics
        • Viral Envelope Proteins / metabolism
        • Viral Matrix Proteins / genetics
        • Viral Matrix Proteins / metabolism

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