Structure, function, and intracellular localization of glycoprotein B of herpesvirus simian agent 8 expressed in insect and mammalian cells.
The cloned gene of glycoprotein B (gB) of herpesvirus simian agent 8 (SA 8) was expressed with a baculovirus system in insect cells. Expression of gB was easily detectable over the cellular background by Coomassie staining of electrophoretically separated proteins. Endoglycosidase digestion of immunoprecipitated gB revealed that the gene product is N-glycosylated, but only with unprocessed, endoglycosidase-H sensitive carbohydrates. The lack of terminal glycosylation of gB is consistent with the observation that gB expressed in insect cells has a molecular weight slightly lower than gB synthesized during an SA 8 infection in mammalian cells. The truncated carbohydrates of gB from insect cells have no measurable effect on the tertiary structure of gB. Immunofluorescence studies on mammalian cells expressing gB from a simian virus 40 based vector revealed that the glycoprotein is localized to cytoplasmic membranes, to the plasma membrane and to the nuclear envelope. Cells expressing gB were fused to polykaryons, which shows that gB has cell fusing activity in the absence of any other SA 8 gene product.
Veit
M
M
Institut für Virologie, Fachbereich Veterinärmedizin, Freie Universität Berlin, Federal Republic of Germany.
Sott
C
C
Borchers
K
K
Ludwig
H
H
Schmidt
M F
MF
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D005822:Genetic Vectors; D006031:Glycosylation; D006568:Herpesvirus 1, Cercopithecine / Q000235:genetics / Q000378:metabolism*; D007425:Intracellular Membranes / Q000737:chemistry; D009036:Moths; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000502:physiology*
D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C069210:SA8 glycoprotein B, simian herpesvirus