Separation of receptor-binding and profusogenic domains of glycoprotein D of herpes simplex virus 1 into distinct interacting proteins.
The 369-residue glycoprotein D (gD) is the entry, receptor-binding protein of herpes simplex virus 1. The common receptors for viral entry are nectin-1, HveA, and a specific O-linked sulfated proteoglycan. The major receptor-binding sites of gD are at the N terminus, whereas the domain required for fusion of viral envelope with the plasma membrane is at the C terminus of the ectodomain (residues 260-310). In the course of retargeting gD to the urokinase plasminogen activator (uPA) receptor for potential therapeutic applications, we obtained a genetically engineered infectious virus in which the receptor-binding domain consisting of the N-terminal domain of uPA fused to residues 33-60 of gD was separated from an independently expressed C-terminal domain of gD containing residues 219-369. The intervening sequences (residues 62-218) were replaced by a stop codon and a promoter for the C-terminal domain of gD. The physical interaction of the two components was reconstructed by coimmunoprecipitation of the N-terminal domain of uPA with the C-terminal domain of gD. These results indicate that codons 61-218 of gD do not encode executable functions required for viral entry into cells and suggest that the receptor-binding ligand must interact with but need not alter the structure of the residual portion of gD to effect virus entry. This finding opens the way for the development of a family of recombinant viruses in which the profusion domain of gD and independently furnished, interacting receptor-binding domains effect entry of the virus via a range of receptors.
Zhou
Guoying
G
The Marjorie B. Kovler Viral Oncology Laboratories, University of Chicago, 910 East 58th Street, Chicago, IL 60637, USA.
Roizman
Bernard
B
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D005818:Genetic Engineering; D018259:Herpesvirus 1, Human / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000737:chemistry*
D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C039447:glycoprotein D, Human herpesvirus 1; D014568:Urokinase-Type Plasminogen Activator