Intracellular transport of recombinant coronavirus spike proteins: implications for virus assembly.
Coronavirus spike protein genes were expressed in vitro by using the recombinant vaccinia virus expression system. Recombinant spike proteins were expressed at the cell surface and induced cell fusion in a host-cell-dependent fashion. The intracellular transport of recombinant spike proteins was studied. The half time of acquisition of resistance to endo-beta-N-acetylglucosaminidase H was approximately 3 h for the recombinant feline infectious peritonitis virus S protein. The S protein in feline infectious peritonitis virus-infected cells was found to have a half time of acquisition of resistance to endo-beta-N-acetylglucosaminidase H of approximately 1 h. This difference can be explained by the fact that coronavirus budding takes place at intracellular membranes and that the oligosaccharides of the spike protein are modified after budding. Apparently, spike protein incorporated into budded virions is transported faster through the Golgi apparatus than is spike protein alone. These findings provide new insights into the mechanism of coronavirus budding and are discussed in relation to current models of intracellular transport and sorting of proteins.
Department of Infectious Diseases and Immunology, Veterinary Faculty, State University, Utrecht, The Netherlands.
H
H
Vennema
L
L
Heijnen
A
A
Zijderveld
M C
MC
Horzinek
W J
WJ
Spaan
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003332:Coronaviridae / Q000235:genetics* / Q000502:physiology; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral; D005822:Genetic Vectors; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D014616:Vaccinia virus / Q000235:genetics; D014764:Viral Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000235:genetics*
D011994:Recombinant Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins