Identification of a site on herpes simplex virus type 1 glycoprotein D that is essential for infectivity.
Herpes simplex virus glycoprotein D (gD) plays an essential role during penetration of the virus into cells. There is evidence that it recognizes a specific receptor after initial attachment of virions to cell surface heparan sulfate and also that gD-1, gD-2, and gp50 (the pseudorabies virus gD homolog) bind to the same receptor. Although the antigenic structure of gD has been studied intensively, little is known about functional regions of the protein. Antigenic site I is a major target for neutralizing antibodies and has been partially mapped by using deletion mutants and neutralization-resistant viruses. Working on the assumption that such a site may overlap with a functional region of gD, we showed previously that combining two or more amino acid substitutions within site I prevents gD-1 from functioning and is therefore lethal. We have now used a complementation assay to measure the functional activity of a panel of deletion mutants and compared the results with an antigenic analysis. Several mutations cause gross changes in protein folding and destroy functional activity, whereas deletions at the N and C termini have little or no effect on either. In contrast, deletion of residues 234 to 244 has only localized effects on antigenicity but completely abolishes functional activity. This region, which is part of antigenic site Ib, is therefore essential for gD-1 function. The complementation assay was also used to show that a gD-negative type 1 virus can be rescued by gD-2 and by two gD-1-gD-2 hybrids but not by gp50, providing some support for the existence of a common receptor for herpes simplex virus types 1 and 2 but not pseudorabies virus. Alternatively, gp50 may lack a signal for incorporation into herpes simplex virions.
Department of Microbiology, University of Pennsylvania, Philadelphia 19104-6003.
M I
MI
Muggeridge
W C
WC
Wilcox
G H
GH
Cohen
R J
RJ
Eisenberg
D000818:Animals; D000911:Antibodies, Monoclonal; D002872:Chromosome Deletion; D000939:Epitopes / Q000032:analysis; D005814:Genes, Viral; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D008961:Models, Structural; D010957:Plasmids; D011487:Protein Conformation; D011991:Receptors, Virus / Q000378:metabolism; D018139:Simplexvirus / Q000235:genetics* / Q000472:pathogenicity; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011991:Receptors, Virus; D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1