Advertisement

Overexpression of TGEV Cell Receptor Impairs the Production of Virus Particles

  • B. Delmas
  • E. Kut
  • J. Gelfi
  • H. Laude
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 380)

Abstract

The porcine aminopeptidase-N (pAPN) is the cellular receptor for the transmissible gastroenteritis virus (TGEV) due to the specific binding of the spike protein S to APN. In the present study, we performed both biological and biochemical experiments to analyze how the level of expression of a virus receptor can influence the viral protein biosynthesis and the virus production. We generated two swine testis cell clones overexpressing pAPN (ST-APN clones). These clones produced 104 less infectious virus than control ST cells. Plaque assays revealed a four-fold reduction of the diameter of the plaques in ST-APN cells compared to ST cells. Pulse-chase experiments revealed that S transport from the endoplasmic reticulum to the Golgi apparatus was not affected in ST-APN cells. Additionally, an anti-APN antibody was able to increase the virus released in the supernatant of ST-APN cells. Likewise, BHK clones expressing variable amounts of pAPN were shown to acquire TGEV susceptibility and to produce infectious particles as an inverse function of their level of pAPN expression. In contrast, MDCK clones expressing low or large amounts of pAPN failed to produce infectious particles. Taken together, these studies strongly suggest that overexpression of receptor, but also other(s) undetermined factor(s), can impair the production of viral particles.

Keywords

MDCK Cell Virus Production Spike Protein Transmissible Gastroenteritis Virus Virus Cycle 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Delmas, B., Gelfi, J., L’Haridon, R., Vogel, L.K., Sjöström, H., Noren, O., Laude, H. Aminopeptidase N is a major receptor for the enteropathogenic coronavirus TGEV. Nature 1992;357:417–420.PubMedCrossRefGoogle Scholar
  2. 2.
    Godet, M., Grosclaude, J., Delmas, B., Laude, H. Major receptor-binding and neutralization determinants are located within the same domain of the coronavirus transmissible gastroenteritis virus spike protein. J.Virol. 1994;68:(in press).Google Scholar
  3. 3.
    Hansen, G., Vogel, L.K., Delmas, B., Gelfi, J., Laude, H., Noren, O., Sjöström, H. The coronavirus TGEV causes infection after endocytosis by an aminopeptidase N recruiting mechanism. (manuscript in preparation).Google Scholar
  4. 4.
    Laude, H., Chapsal, J.-M., Gelfi, J., Labiau, S., Grosclaude, J. Antigenic structure of transmissible gastroenteritis virus. I. Properties of monoclonal antibodies directed against virion proteins. 1986; 67:119–130.Google Scholar
  5. 5.
    Delmas, B., Laude, H. Assembly of coronavirus spike protein into trimers and its role in epitope expression. J. Virol. 1990;64:5367–5375.PubMedGoogle Scholar
  6. 6.
    Louvard, D. Apical membrane aminopeptidase appears at site of cell-cell contact in cultured kidney epithelial cells. Proc. Natl. Acad. Sci. USA 1980;77:4132–4136.PubMedCrossRefGoogle Scholar
  7. 7.
    Pensaert, M., Callebaut, P., Cox, E. Enteric coronaviruses of animals. In: Kapikian A.Z., (ed.) Viral infections of the gastrointestinal tract. 2nd ed. Marcel Dekker, Inc., New York 1993 pp 627-696.Google Scholar
  8. 8.
    Bour, S., Geleziunas, R., Wainberg, M.A. The role of CD4 and its downmodulation in establisment and maintenance of HIV-1 infection. Immunological Reviews. 1994;140:147–171.PubMedCrossRefGoogle Scholar
  9. 9.
    Naniche, D., Wild, T.F., Rabourdin-Combe, C., Gerlier, D. Measles virus haemagglutinin induces down-regulation of gp57/67, a molecule involved in virus binding. J. Gen. Virol. 1993;74:1073–1079.PubMedCrossRefGoogle Scholar
  10. 10.
    Delmas, B., Gelfi, J., Kut, E., Sjöström, H., Noren, O., Laude, H. Determinants essential for the transmissible gastroenteritis virus-receptor interaction reside within a domain of aminopeptidase-N that is distinct from the enzymatic site. J. Virol. 1994;68:5216–5224.PubMedGoogle Scholar
  11. 11.
    Kreutz, L.C., Seal, B.S., Mengeling, W.L. Early interaction of feline calicivirus with cells in culture. Arch. Virol. 1994;136:19–34.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • B. Delmas
    • 1
  • E. Kut
    • 1
  • J. Gelfi
    • 1
  • H. Laude
    • 1
  1. 1.Unite de Virologie et Immunologie MoléculairesINRAJouy-en-JosasFrance

Personalised recommendations