Influence of new glycosylation sites on expression of the vesicular stomatitis virus G protein at the plasma membrane.
In this report, we have asked whether asparagine-linked oligosaccharides added to new sites in the polypeptide backbone of a model plasma membrane glycoprotein, the vesicular stomatitis virus G protein, can promote its intracellular transport. We modified the coding sequence of G protein lacking the two normal consensus sites for glycosylation by oligonucleotide-directed mutagenesis to create new consensus sites. The expression of the mutant proteins was then analyzed in transfected cells. Six of the eight new sites which were introduced were glycosylated, and an oligosaccharide at two of these new sites promoted transport of G protein which lacked the two normal sites. However, the efficiency of this process was reduced compared to the wild-type protein or to the proteins with only one oligosaccharide at either of the normal sites. In addition, an oligosaccharide at two of the other new sites caused inhibition of transport of the wild-type G protein. The data in this and the following report suggest that carbohydrate plays an indirect role in the intracellular transport of G protein.
Machamer
C E
CE
Molecular Biology and Virology Laboratory, Institute for Biological Studies, San Diego, California 92138.
Rose
J K
JK
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D005455:Fluorescent Antibody Technique; D006031:Glycosylation; D007163:Immunosorbent Techniques; D007457:Iodine Radioisotopes / Q000378:metabolism; D007784:Lactoperoxidase / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D009844:Oligosaccharides / Q000378:metabolism; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology; D014721:Vesicular stomatitis Indiana virus; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins / Q000235:genetics / Q000378:metabolism*
D004274:DNA, Recombinant; C028391:G protein, vesicular stomatitis virus; D007457:Iodine Radioisotopes; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; D014415:Tunicamycin; D007784:Lactoperoxidase