The cytoplasmic domain of herpes simplex virus type 1 glycoprotein C is required for membrane anchoring.
The herpes simplex virus type 1 (HSV-1) glycoprotein C (gC) gene was altered so that it encoded a truncated glycoprotein lacking a cytoplasmic domain but retaining 20 of 23 amino acids of the transmembrane domain. No additional amino acid residues were introduced into the glycoprotein encoded by the altered gene. The gene was recombined into the HSV-1 genome by marker transfer. Two recombinant viruses, dl1 and dl2, that expressed the mutant gene were isolated. Characterization of these viruses showed that a substantial fraction of the mutant glycoprotein was secreted from infected cells. Pulse-chase experiments showed that the kinetics of posttranslational modification of the mutant glycoprotein were similar to those of the wild type. However, comparison of the kinetics of secretion of gC by dl2 and gC-3, a gC mutant lacking both the transmembrane and cytoplasmic domains, showed that dl2 gC was secreted much more slowly than gC-3 gC. Iodination of plasma membrane glycoproteins showed that dl2 gC was initially expressed on the cell surface as a membrane protein and subsequently was slowly released from the membrane into the medium. These data indicate that a major function of the cytoplasmic domain of gC is to ensure the stable anchoring of the glycoprotein in plasma membranes. In contrast to these major changes in the membrane-anchoring properties of gC, characterization of the virions produced by dl1 and dl2 showed that they contain significant amounts of gC. Thus the cytoplasmic domain does not appear to be essential for incorporation of this glycoprotein into virions.
Holland
T C
TC
Department of Immunology and Microbiology, Wayne State University Medical School, Detroit, Michigan 48201.
Lerch
R J
RJ
Earhart
K
K
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005838:Genotype; D006801:Humans; D009154:Mutation; D010957:Plasmids; D013329:Structure-Activity Relationship; D014759:Viral Envelope Proteins / Q000032:analysis* / Q000235:genetics
D014759:Viral Envelope Proteins; C038535:glycoprotein gC, herpes simplex virus type 1