Expression of Semliki Forest virus proteins from cloned complementary DNA. I. The fusion activity of the spike glycoprotein.
A complementary (cDNA) molecule encoding the structural proteins of Semliki Forest virus (SFV) has been inserted into a Simian virus 40-derived eucaryotic expression vector lacking introns. Introduction of the recombinant DNA into nuclei of baby hamster kidney cells results in the synthesis of authentic SFV membrane glycoproteins E1 and E2. The glycoproteins are both transported to the cell surface and induce cell-cell fusion after a brief treatment of the cells with low pH medium. The pH dependence of the fusion reaction was the same as that induced by virus particles (White, J., J. Kartenbeck, and A. Helenius, 1980, J. Cell Biol., 89:674-679). Transfection of cells with another recombinant DNA molecule in which the SFV cDNA is engineered into the same expression vector including an intron has been shown before to result in the expression of only the E2 protein on the cell surface, whereas the E1 protein is trapped in the rough endoplasmic reticulum (Kondor-Koch, C., H. Riedel, K. Söderberg, and H. Garoff, 1982, Proc. Natl. Acad. Sci. USA, 79:4525-4529). Such cells do not exhibit pH-dependent polykaryon formation, suggesting that the E1 protein is necessary for fusion activity. Immunoblotting experiments show that the RER-trapped E1 protein expressed from the DNA construction with an intron has a smaller apparent molecular weight than authentic E1, and that is has lost its amphipathic characteristics.
D000818:Animals; D002451:Cell Compartmentation; D002478:Cells, Cultured; D006224:Cricetinae; D004247:DNA / Q000235:genetics; D005786:Gene Expression Regulation; D006863:Hydrogen-Ion Concentration; D008561:Membrane Fusion; D008565:Membrane Proteins / Q000235:genetics*; D010957:Plasmids; D012672:Semliki forest virus; D014760:Viral Fusion Proteins; D014764:Viral Proteins / Q000235:genetics*
D008565:Membrane Proteins; D014760:Viral Fusion Proteins; D014764:Viral Proteins; D004247:DNA