The small envelope glycoprotein (GS) of equine arteritis virus folds into three distinct monomers and a disulfide-linked dimer.
The small membrane glycoprotein (GS) of equine arteritis virus (EAV) is a minor virion component but is abundantly expressed in EAV-infected cells. In this study, we have analyzed its membrane topology, folding, oligomerization, and intracellular transport. We show that GS is a class I integral membrane protein with one functional N-glycosylation site. Gel electrophoresis under nonreducing conditions revealed that GS occurs in EAV-infected cells in four monomeric conformations and as disulfide-linked homodimers. The slowest-migrating monomeric form corresponded to the fully reduced GS protein; the three faster-migrating monomeric species are probably generated by the formation of alternative intrachain disulfide bonds between the three luminal cysteines in the molecule. The GS monomers were selectively retained in the endoplasmic reticulum, as judged by their permanent susceptibility to endoglycosidase H, whereas the GS dimers were specifically incorporated into virus particles and became endoglycosidase H resistant and sialylated during passage through the Golgi apparatus.
de Vries
A A
AA
Department of Infectious Diseases and Immunology, Veterinary Faculty, Utrecht University, The Netherlands.
Raamsman
M J
MJ
van Dijk
H A
HA
Horzinek
M C
MC
Rottier
P J
PJ
D000595:Amino Acid Sequence; D000818:Animals; D001704:Biopolymers; D002460:Cell Line; D006224:Cricetinae; D004220:Disulfides / Q000378:metabolism*; D007240:Equartevirus / Q000378:metabolism*; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017510:Protein Folding; D011499:Protein Processing, Post-Translational; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D001704:Biopolymers; D004220:Disulfides; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; C093519:glycoprotein S, equine arteritis virus