title: Mutational analysis of the role of glycoprotein I in varicella-zoster virus replication and its effects on glycoprotein E conformation and trafficking.
abstract: The contributions of the glycoproteins gI (ORF67) and gE (ORF68) to varicella-zoster virus (VZV) replication were investigated in deletion mutants made by using cosmids with VZV DNA derived from the Oka strain. Deletion of both gI and gE prevented virus replication. Complete deletion of gI or deletions of 60% of the N terminus or 40% of the C terminus of gI resulted in a small plaque phenotype as well as reduced yields of infectious virus. Melanoma cells infected with gI deletion mutants formed abnormal polykaryocytes with a disrupted organization of nuclei. In the absence of intact gI, gE became localized in patches on the cell membrane, as demonstrated by confocal microscopy. A truncated N-terminal form of gI was transported to the cell surface, but its expression did not restore plaque morphology or infectivity. The fusogenic function of gH did not compensate for gI deletion or the associated disruption of the gE-gI complex. These experiments demonstrated that gI was dispensable for VZV replication in vitro, whereas gE appeared to be required. Although VZV gI was dispensable, its deletion or mutation resulted in a significant decrease in infectious virus yields, disrupted syncytium formation, and altered the conformation and distribution of gE in infected cells. Normal cell-to-cell spread and replication kinetics were restored when gI was expressed from a nonnative locus in the VZV genome. The expression of intact gI, the ORF67 gene product, is required for efficient membrane fusion during VZV replication.
authors.affiliation: Department of Pediatrics and Microbiology/Immunology, Stanford University School of Medicine, California 94305, USA.
authors.forename: S
authors.initials: S
authors.lastname: Mallory
authors.forename: M
authors.initials: M
authors.lastname: Sommer
authors.forename: A M
authors.initials: AM
authors.lastname: Arvin
mesh_terms: D001692:Biological Transport; D015153:Blotting, Western; D002459:Cell Fusion; D002462:Cell Membrane / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D014645:Herpesvirus 3, Human / Q000254:growth & development*; D016254:Mutagenesis, Insertional; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D017384:Sequence Deletion; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins / Q000502:physiology* / Q000648:ultrastructure; D014779:Virus Replication
chemical_list: D014759:Viral Envelope Proteins; C056913:glycoprotein E, varicella-zoster virus; C070916:glycoprotein IV, varicella-zoster virus