Cell surface expression of herpes simplex virus type 1 glycoprotein H in recombinant baculovirus-infected cells.
DNA encoding the complete sequence for the herpes simplex virus type 1 (HSV-1) glycoprotein H (gH) was inserted into a baculovirus transfer vector under control of the baculovirus polyhedrin gene promoter. After co-transfection with wild-type baculovirus DNA, recombinants expressing gH were isolated by plaque purification. The baculovirus-expressed HSV-1 gH represented a significant portion of total cellular protein and was several hundred fold more abundant than gH in HSV-1-infected Vero cells. The expressed gH appeared to be glycosylated, since it was similar in size to wild-type HSV-1 gH, was susceptible to both tunicamycin and endoglycosidase-H treatment, and was labeled by [3H]mannose. In contrast to previous reports of gH expressed in mammalian cells, the baculovirus recombinant-expressed gH was abundant on the cell surface as judged by indirect immunofluorescence. To our knowledge, this is the first report of expressed HSV-1 gH being transported to the cell surface in the absence of other HSV-1 gene products and the first report of expressed gH with an apparent molecular weight similar to authentic HSV-1 gH.
Ghiasi
H
H
Ophthalmology Research, Cedars-Sinai Medical Center, Los Angeles, California 90048.
Nesburn
A B
AB
Wechsler
S L
SL
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D002462:Cell Membrane / Q000648:ultrastructure; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors; D007313:Insecta; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000494:pharmacology; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D015183:Restriction Mapping; D018139:Simplexvirus / Q000235:genetics*; D014162:Transfection; D014415:Tunicamycin / Q000494:pharmacology; D014759:Viral Envelope Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*
D004279:DNA, Viral; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C044200:glycoprotein H, herpes simplex virus type 1; D014415:Tunicamycin; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase