Glycoprotein B of human herpesvirus 8 is a component of the virion in a cleaved form composed of amino- and carboxyl-terminal fragments.
Human herpesvirus 8 (HHV-8) or Kaposi's sarcoma-associated herpesvirus (KSHV) is the only known human member of the Rhadinovirus genus of the gammaherpesvirus subfamily. Antibodies against peptides representing portions of the amino- and carboxyl-termini of HHV-8 gB were produced and used to detect gB expression in Vero cells transfected with the gB gene, in the HHV-8-harboring cell line, BCBL-1, and in purified virions. Expression of gB was detected in approximately 3% of uninduced BCBL-1 cells, while up to 30% of the cells expressed gB after 12-O-tetradecanoylphorbol-13-acetate (TPA) induction of virus replication. Indirect immunofluorescence assays and confocal microscopy showed that gB was distributed throughout the cytoplasm of BCBL-1 cells and transfected Vero cells. Immunoblot analyses of virion preparations revealed the presence of full-length as well as two smaller than full-length gB-derived species corresponding to the amino- and carboxy-terminal portions of gB, respectively. Biochemical analysis of the gB carbohydrate moieties using glycosylation inhibitors revealed that gB contained N-linked oligosaccharides of the high-mannose type, characteristic of precursor carbohydrate chains added in the endoplasmic reticulum.
Baghian
A
A
Department of Veterinary Microbiology and Parasitology, School of Veterinary Medicine, Baton Rouge, LA 70803, USA.
Luftig
M
M
Black
J B
JB
Meng
Y X
YX
Pau
C P
CP
Voss
T
T
Pellett
P E
PE
Kousoulas
K G
KG
D000581:Amidohydrolases / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000821:virology; D019084:Fluorescent Antibody Technique, Indirect; D006031:Glycosylation / Q000187:drug effects; D019288:Herpesvirus 8, Human / Q000737:chemistry* / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism; D006596:Hexosaminidases / Q000378:metabolism; D008358:Mannose / Q000378:metabolism; D008970:Molecular Weight; D010446:Peptide Fragments / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D011499:Protein Processing, Post-Translational; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured; D014415:Tunicamycin / Q000494:pharmacology; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects
D010446:Peptide Fragments; D014759:Viral Envelope Proteins; C116207:glycoprotein B, human herpesvirus 8; D014415:Tunicamycin; D006596:Hexosaminidases; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D013755:Tetradecanoylphorbol Acetate; D008358:Mannose