The rubella virus E2 and E1 spike glycoproteins are targeted to the Golgi complex.
Rubella virus (RV) has been reported to bud from intracellular membranes in certain cell types. In this study the intracellular site of targeting of RV envelope E2 and E1 glycoproteins has been investigated in three different cell types (CHO, BHK-21 and Vero cells) transfected with a cDNA encoding the two glycoproteins. By indirect immunofluorescence, E2 and E1 were localized to the Golgi region of all three cell types, and their distribution was disrupted by treatment with BFA or nocodazole. Immunogold labeling demonstrated that E2 and E1 were localized to Golgi cisternae and indicated that the glycoproteins were distributed across the Golgi stack. Analysis of immunoprecipitates obtained from stably transfected CHO cells revealed that E2 and E1 become endo H resistant and undergo sialylation without being transported to the cell surface. Transport of RV glycoproteins to the Golgi complex was relatively slow (t1/2 = 60-90 min). Coprecipitation experiments indicated that E2 and E1 form a heterodimer in the RER. E1 was found to fold much more slowly than E2, suggesting that the delay in transport of the heterodimer to the Golgi may be due to the slow maturation of E1 in the ER. These results indicate that RV glycoproteins behave as integral membrane proteins of the Golgi complex and thus provide a useful model to study targeting and turnover of type I membrane proteins in this organelle.
Hobman
T C
TC
Division of Cellular and Molecular Medicine, University of California, San Diego, La Jolla 92093-0651.
Woodward
L
L
Farquhar
M G
MG
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis* / Q000378:metabolism; D020126:Brefeldin A; D016466:CHO Cells / Q000737:chemistry / Q000378:metabolism; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003517:Cyclopentanes / Q000494:pharmacology; D006023:Glycoproteins / Q000032:analysis / Q000378:metabolism; D006056:Golgi Apparatus / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D008954:Models, Biological; D015739:Nocodazole / Q000494:pharmacology; D012412:Rubella virus / Q000378:metabolism*; D014709:Vero Cells / Q000737:chemistry / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000032:analysis* / Q000378:metabolism
D000956:Antigens, Viral; D003517:Cyclopentanes; D006023:Glycoproteins; D014759:Viral Envelope Proteins; C061826:E1 envelope protein, Rubella virus; D020126:Brefeldin A; D015739:Nocodazole