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.
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References
Armstrong JH, Niemann H, Smeekens S, Rottier P, Warren G (1984) Sequence and topology of a model intracellular membrane protein, El glycoprotein, from a coronavirus. Nature 308: 751–752
Braakman I, Hoover-Litty H, Wagner KR, Helenius A (1991) Folding of influenza hemagglutinin in the endoplasmic reticulum. J Cell Biol 114: 401–411
Braakman I, Helenius J, Helenius A (1992) Manipulating disulfide bond formation and protein folding in the endoplasmic reticulum. EMBO J 11: 1717–1722
Griffiths G, Rottier P (1992) Cell biology of viruses that assemble along the biosynthetic pathway. Semin Cell Biol 3: 367–381
Holmes KV, Doller EW, Sturman LS (1981) Tunicamycin resistant glycosylation of a coronavirus glycoprotein: demonstration of a novel type of viral glycoprotein. Virology 115: 334–344
Hurtley SM, Helenius A (1989) Protein oligomerization in the endoplasmic reticulum. Annu Rev Cell Biol 5: 277–307
Krijnse Locker J, Griffiths G, Horzinek MC, Rottier PJM (1992) O-glycosylation of the Coronavirus M protein: differential localization of sialyltransferases in N- and O-linked glycosylation. J Biol Chem 267: 14094–14101
Pettersson R (1991) Protein localization and virus assembly at intracellular membranes. In: Compans RW (ed) Protein traffic in eukaryotic cells. Curr Top Microbiol Immunol 170: 67–106
Rottier PJM, Horzinek MC, van der Zeijst BAM (1981) Viral protein synthesis in mouse hepatitis virus strain A59-infected cells: effects of tunicamycin. J Virol 40: 350–357
Rottier PJM, Welling GW, Welling-Wester S, Niesters HGM, Lenstra JA, van der Zeijst BAM (1986) Predicted membrane topology of the Coronavirus protein El. Biochemistry 25: 1335–1339
Rottier PJM, Rose JK (1987) Coronavirus E1 glycoprotein expressed from cloned cDNA localizes in the Golgi region. J Virol 61: 2042–2045
Simons K, Garoff H (1980) The budding mechanism of enveloped animal viruses. J Gen Virol 50: 1–21
Tooze J, Tooze S, Warren G (1984) Replication of Coronavirus MHV-A59 in sac- cells: determination of the first site of budding of progeny virions. Eur J Cell Biol 33: 281–293
Tooze J, Tooze S, Warren G (1988) Site of addition of N-acetylgalactosamine to the E1 glycoprotein of mouse hepatitis virus-A59. J Cell Biol 106: 1475–1487
Vennema H, Rottier PJM, Heijnen L, Godeke GJ, Horzinek MC, Spaan WJM (1990) Biosynthesis and function of the Coronavirus spike protein. In: Cavanagh D, Brown TDK (eds) Coronaviruses and their diseases. Plenum Press, New York, pp 9–19
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© 1994 Springer-Verlag
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Opstelten, DJ.E., de Groote, P., Horzinek, M.C., Rottier, P.J.M. (1994). Folding of the mouse hepatitis virus spike protein and its association with the membrane protein. In: Brinton, M.A., Calisher, C.H., Rueckert, R. (eds) Positive-Strand RNA Viruses. Archives of Virology Supplementum, vol 9. Springer, Vienna. https://doi.org/10.1007/978-3-7091-9326-6_32
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DOI: https://doi.org/10.1007/978-3-7091-9326-6_32
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