Processing of human cytomegalovirus envelope glycoproteins in and egress of cytomegalovirus from human astrocytoma cells.
The synthesis of human cytomegalovirus (HCMV) envelope glycoproteins and the production of infectious HCMV in human astrocytoma and skin fibroblast (SF) cells were analysed. HCMV envelope glycoproteins synthesized in astrocytoma cells had lower Mrs than the same glycoproteins synthesized in SF cells regardless of the strain of HCMV used, showing that the differences observed were due to differences in processing by the host cell and not the strain of HCMV used. HCMV envelope glycoproteins synthesized in astrocytoma cells were found to contain less galactosamine. Moreover, when synthesized in SF cells some HCMV glycoproteins contained a protease-resistant fragment owing to the presence of a cluster of O-linked oligosaccharides on the polypeptide. This fragment was not present when these HCMV glycoproteins were synthesized in astrocytoma cells. These data suggested that HCMV glycoproteins synthesized in astrocytoma cells contain fewer O-linked oligosaccharides. In contrast, other post-translational events such as proteolytic cleavage of the HCMV gB glycoprotein and the formation of disulphide-linked complexes did occur. The virus produced in astrocytoma cells was capable of infecting SF cells, suggesting that complete O-glycosylation is not needed to produce infectious HCMV. However, astrocytoma cells were slow to release virus into the culture medium, suggesting that a fully functional Golgi network is needed for efficient egress of HCMV from the host cell.
Kari
B
B
Biomedical Research Institute, Children's Hospital, St Paul, Minnesota 55102.
Radeke
R
R
Gehrz
R
R
D001254:Astrocytoma; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003587:Cytomegalovirus / Q000254:growth & development / Q000502:physiology*; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000382:microbiology; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000096:biosynthesis / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism / Q000382:microbiology*; D006801:Humans; D008970:Molecular Weight; D011233:Precipitin Tests; D012867:Skin / Q000166:cytology; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000378:metabolism*; D010948:Viral Plaque Assay
D006023:Glycoproteins; D014759:Viral Envelope Proteins; C073092:cytomegalovirus glycocomplex I; C073093:cytomegalovirus glycocomplex II; C040495:glycoprotein B, Simplexvirus