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Folding of the mouse hepatitis virus spike protein and its association with the membrane protein

  • D.-J. E. Opstelten
  • P. de Groote
  • M. C. Horzinek
  • P. J. M. Rottier
Part of the Archives of Virology Supplementum book series (ARCHIVES SUPPL, volume 9)

Summary

Coronaviruses are assembled by budding into pre-Golgi membranes. Using different approaches we have demonstrated that the spike (S) protein and the membrane (M) protein of mouse hepatitis virus (MHV) associate to form large complexes. Newly synthesized M was found in these complexes almost immediately after its synthesis, whereas the S protein started to appear in heterocomplexes after 10–20 min. This is consistent with the slow rate of folding of S and with the observation that folding of S preceeds its association with M. While the folding of S involves the formation of multiple disulfide bonds, folding of M is disulfide-independent. This contrast was reflected by the differential sensitivity of the two proteins to reduction with dithiothreitol (DTT). Addition of DTT to the culture medium of MHV-infected cells drastically impaired the folding of S, but not of M. Consequently, the S protein was unable to interact with M. Under these conditions, S stayed in the ER while M was transported efficiently beyond the site of budding to the Golgi complex. We conclude that the association of S with M is an essential step in the formation of the viral envelope and in the accumulation of both proteins at the site of virus assembly.

Keywords

Envelope Protein Golgi Complex Nucleocapsid Protein Disulfide Bond Formation Mouse Hepatitis Virus 
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.

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Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • D.-J. E. Opstelten
    • 1
  • P. de Groote
    • 1
  • M. C. Horzinek
    • 1
  • P. J. M. Rottier
    • 1
  1. 1.Institute of Virology, Department of Infectious Diseases and Immunology, Veterinary FacultyUtrecht UniversityThe Netherlands

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