Deletions in herpes simplex virus glycoprotein D define nonessential and essential domains.
Herpes simplex virus glycoprotein D (gD) is a major component of the virion envelope and infected cell membranes and is essential for virus entry into cells. We have recently shown that gD interacts with a limited number of cell surface receptors which are required for virus penetration into cells. To define domains of gD which are required for aspects of virus replication including receptor binding, deletion mutations of 5 to 14 amino acids were constructed by using oligonucleotide-directed mutagenesis. Plasmids containing mutant genes for gD were assayed for the ability to rescue a recombinant virus, F-gD beta, in which beta-galactosidase sequences replace gD-coding sequences. Effects on global folding and posttranslational processing of the molecules were assessed by using a panel of monoclonal antibodies which recognize both continuous and discontinuous epitopes. A region near the amino terminus (residues 7 to 21) of gD which is recognized by monoclonal antibodies able to neutralize herpes simplex virus in the absence of complement was not essential for function. In addition, virtually all of the cytoplasmic domain of gD and an extracellular domain close to the membrane were dispensable. In contrast, deletion mutations in the central region of the molecule, save for one exception, led to alterations in global folding of the molecule and maturation of the protein was inhibited.
Feenstra
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V
Department of Pathology, McMaster University, Hamilton, Ontario, Canada.
Hodaie
M
M
Johnson
D C
DC
D000818:Animals; D002460:Cell Line; D002872:Chromosome Deletion; D015870:Gene Expression; D005814:Genes, Viral; D006801:Humans; D009154:Mutation; D010957:Plasmids; D018139:Simplexvirus / Q000235:genetics* / Q000648:ultrastructure; D014162:Transfection; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000235:genetics*; D010948:Viral Plaque Assay
D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1