Comparison of rabies virus G proteins produced by cDNA-transfected animal cells that display either inducible or constitutive expression of the gene.
By using a retrovirus expression vector, pZIP-NeoSV(X)1, we introduced a cloned cDNA of the rabies virus G gene into BHK-21 cells and the NA cell clone originated from the murine neuroblastoma C1300 line. Using the neomycin resistance gene of the vector, we isolated several G418-resistant transformants of BHK-21 and NA cells (referred to as G-BHK and G-NA cells, respectively). G-BHK cells constitutively produced G proteins, whereas G-NA cells produced the proteins only when treated with sodium butyrate. G proteins synthesized in these transformants were transported normally to the surface of the cell, but they displayed different electrophoretic mobilities, which were shown to originate from differences in the number and structure of the carbohydrate moieties of the protein; G-BHK cells produced highly glycosylated and sialylated G proteins, whereas less glycosylated and much less sialylated G proteins were produced by G-NA cells as observed in virus-infected NA and BHK-21 cells, indicating that the glycosylation and sialylation of the G protein depend on the cellular conditions under which the protein was produced. In the absence of sodium butyrate the G protein was not detectable in G-NA cells either by immunoblot assay or fluorescent antibody staining, but the cells were fairly sensitive to syngeneic rabies virus-specific cytotoxic T lymphocytes, although the sensitivity was much increased by treatment with sodium butyrate.
Morimoto
K
K
Department of Molecular Microbiology, Faculty of Pharmaceutical Sciences, Kyoto University, Japan.
Kawai
A
A
Mifune
K
K
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D002460:Cell Line; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D005455:Fluorescent Antibody Technique; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D015151:Immunoblotting; D011233:Precipitin Tests; D011820:Rabies virus / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014162:Transfection; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics
D000956:Antigens, Viral; D002087:Butyrates; D004279:DNA, Viral; D006023:Glycoproteins; D014759:Viral Envelope Proteins; C064951:glycoprotein G, Rabies virus; D020148:Butyric Acid