title: Analysis of the intracellular maturation of the herpes simplex virus type 1 glycoprotein gH in infected and transfected cells.
abstract: We have expressed the HSV-1 glycoprotein, gH, in transiently transfected COS-1 cells. The expressed protein was retained intracellularly, contained unprocessed carbohydrate, and was unrecognized by the monoclonal antibody, LP11. In addition, the protein was aggregated. These properties suggest that unlike other HSV glycoproteins, gH is misfolded in transfected cells. Pulse-chase studies of HSV-1-infected cells indicate that the kinetics of processing of gH are comparable to those of gB, gC, and gD. Rescue studies suggest that gH may interact with another protein during maturation in infected cells. However, we were unable to detect any stable interaction, although analysis of gH on neutral sucrose gradients shortly after synthesis indicated a possible transient association with a high molecular weight molecule or complex. The processing and cell surface expression of gH were also analyzed in HSV-1 virus mutants lacking gB, gC, or gD. Our results indicate that the maturation and cell surface transport of gH did not require the presence of these HSV-1 glycoproteins. In addition, three truncation mutants were constructed by linker insertion mutagenesis. Each of the three truncated proteins was synthesized, but the proteins were aggregated, contained only endo H-sensitive carbohydrate, and none were secreted.
authors.affiliation: Department of Microbiology, School of Dental Medicine, University of Pennsylvania, Philadelphia 19104-6003.
authors.forename: S R
authors.initials: SR
authors.lastname: Roberts
authors.forename: M
authors.initials: M
authors.lastname: Ponce de Leon
authors.forename: G H
authors.initials: GH
authors.lastname: Cohen
authors.forename: R J
authors.initials: RJ
authors.lastname: Eisenberg
mesh_terms: D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000276:immunology / Q000378:metabolism; D001692:Biological Transport; D015153:Blotting, Western; D005814:Genes, Viral; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006031:Glycosylation; D006596:Hexosaminidases / Q000494:pharmacology; D046911:Macromolecular Substances; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008562:Membrane Glycoproteins / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011485:Protein Binding; D011499:Protein Processing, Post-Translational; D018139:Simplexvirus / Q000235:genetics*; D013329:Structure-Activity Relationship; D014162:Transfection; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000235:genetics
chemical_list: D000956:Antigens, Viral; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D015678:Viral Structural Proteins; C044200:glycoprotein H, herpes simplex virus type 1; D006026:Glycoside Hydrolases; D006596:Hexosaminidases; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase