Document Detail

Stable expression of rabies virus glycoprotein in Chinese hamster ovary cells.
MedLine Citation:
PMID:  1993876     Owner:  NLM     Status:  MEDLINE    
The rabies virus glycoprotein (G protein) has several important functions and is a major antigenic stimulus of the host immune system following rabies virus infection or vaccination. We developed a model system for studying the role of N-linked glycosylation in the intracellular transport and antigenicity of this molecule. The full-length cDNA of the G protein of the ERA strain of rabies virus was inserted into the eukaryotic shuttle vector pSG5 and then stably transfected into wild-type Chinese hamster ovary (CHO) cells and mutant CHO cell lines defective in glycosylation. Transfected wild-type CHO cells expressed the G protein (detected by immunofluorescence) on the cell surface in a manner similar to rabies virus-infected cells. The transfected wild-type CHO cells were shown by immunoprecipitation to produce a protein of 67K that comigrated with the fully glycosylated G protein isolated from virus-infected cells or purified virions. Treatment of the transfected cell lines with tunicamycin completely blocked surface expression and resulted in the intracellular accumulation of the G protein, suggesting that the presence of N-linked oligosaccharides is important for transport of this glycoprotein to the plasma membrane. The G protein cDNA was also expressed in the lectin-resistant CHO cell lines Lec 1, Lec 2 and Lec 8. In these cells initial N-linked glycosylation does occur, but later steps in processing of the oligosaccharides are blocked. In each case, the G protein was expressed on the surface of lectin-resistant CHO cells in a similar manner to expression on wild-type CHO cells. This suggests that various different N-linked oligosaccharide structures support intracellular transport of this glycoprotein. Thus, stably transfected CHO cell lines will provide a useful model system for further studies of the role of N-linked glycosylation in trafficking and antigenicity of the rabies virus G protein.
S R Burger; A T Remaley; J M Danley; J Moore; R J Muschel; W H Wunner; S L Spitalnik
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The Journal of general virology     Volume:  72 ( Pt 2)     ISSN:  0022-1317     ISO Abbreviation:  J. Gen. Virol.     Publication Date:  1991 Feb 
Date Detail:
Created Date:  1991-03-21     Completed Date:  1991-03-21     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0077340     Medline TA:  J Gen Virol     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  359-67     Citation Subset:  IM    
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia.
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MeSH Terms
Antigens, Viral / immunology
Biological Transport
Blotting, Southern
Carbohydrate Sequence
Cell Line
Fluorescent Antibody Technique
Gene Expression
Glycoproteins / genetics*,  immunology,  metabolism
Molecular Sequence Data
Precipitin Tests
Rabies virus / genetics*
Tunicamycin / pharmacology
Viral Envelope Proteins / genetics*,  immunology,  metabolism
Grant Support
Reg. No./Substance:
0/Antigens, Viral; 0/Glycoproteins; 0/Lectins; 0/Viral Envelope Proteins; 11089-65-9/Tunicamycin

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