Role of N-linked oligosaccharides in processing and intracellular transport of E2 glycoprotein of rubella virus.
The role of N-linked glycosylation in processing and intracellular transport of rubella virus glycoprotein E2 has been studied by expressing glycosylation mutants of E2 in COS cells. A panel of E2 glycosylation mutants were generated by oligonucleotide-directed mutagenesis. Each of the three potential N-linked glycosylation sites was eliminated separately as well as in combination with the other two sites. Expression of the E2 mutant proteins in COS cells indicated that in rubella virus M33 strain, all three sites are used for the addition of N-linked oligosaccharides. Removal of any of the glycosylation sites resulted in slower glycan processing, lower stability, and aberrant disulfide bonding of the mutant proteins, with the severity of defect depending on the number of deleted carbohydrate sites. The mutant proteins were transported to the endoplasmic reticulum and Golgi complex but were not detected on the cell surface. However, the secretion of the anchor-free form of E2 into the medium was not completely blocked by the removal of any one of its glycosylation sites. This effect was dependent on the position of the deleted glycosylation site.
Qiu
Z
Z
Department of Pathology, University of British Columbia, Vancouver, Canada.
Hobman
T C
TC
McDonald
H L
HL
Seto
N O
NO
Gillam
S
S
D000818:Animals; D001483:Base Sequence; D001693:Biological Transport, Active; D002478:Cells, Cultured; D004252:DNA Mutational Analysis; D004220:Disulfides / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006031:Glycosylation; D006651:Histocytochemistry; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D012412:Rubella virus / Q000737:chemistry / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*
D004220:Disulfides; D009844:Oligosaccharides; D011134:Polysaccharides; D014759:Viral Envelope Proteins; C061827:E2 envelope protein, Rubella virus; D006026:Glycoside Hydrolases