Cell surface expression of a functional rubella virus E1 glycoprotein by addition of a GPI anchor.
Rubella virus (RV) envelope glycoproteins E1 and E2 are targeted to the Golgi as heterodimers. While E2 contains a transmembrane Golgi retention signal, E1 is arrested in a pre-Golgi compartment in the absence of E2, and appears to require heterodimerization in order to reach the Golgi. Various forms of E1 with deletions in the ectodomain or lacking the cytoplasmic (CT) and transmembrane (TM) domains, as well as the 29 C-terminal amino acid residues of the ectodomain were also retained intracellularly. We therefore investigated the possibility of targetting E1 to the plasma membrane by addition of a glycosylphosphatidylinositol (GPI) anchor. We found that E1GPI was transported to the cell surface where it retained the hemadsorption activity characteristic of the wild-type E1/E2 heterodimer. Furthermore, coexpression of a mammalian GPI-specific phospholipase D (GPI-PLD) resulted in the release of E1GPI and in constitutive expression of a soluble form of E1. This study thus demonstrates that the GPI anchor has a dominant effect over the E1 pre-Golgi retention signal and that E1 is sufficient for hemadsorption.
Bernasconi
E
E
Institut de Biologie Animale, Université de Lausanne, Switzerland.
Fasel
N
N
Wittek
R
R
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism / Q000821:virology; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D017261:Glycosylphosphatidylinositols / Q000737:chemistry / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism / Q000821:virology; D006382:Hemadsorption; D006387:Hemagglutination, Viral; D008969:Molecular Sequence Data; D010739:Phospholipase D / Q000235:genetics / Q000378:metabolism; D021382:Protein Sorting Signals / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012412:Rubella virus / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D018076:DNA, Complementary; D004279:DNA, Viral; D017261:Glycosylphosphatidylinositols; D021382:Protein Sorting Signals; D014759:Viral Envelope Proteins; C061826:E1 envelope protein, Rubella virus; D010739:Phospholipase D; C065017:glycoprotein phospholipase D