Random Mutagenesis of the Gene Encoding a Viral Ligand for Multiple Cell Entry Receptors to Obtain Viral Mutants Altered for Receptor Usage

Herpes simplex virus type 1 (HSV-1) can enter cells expressing any one of multiple entry receptors, including the herpesvirus entry mediator (HVEM), nectin-1, and sites in heparan sulfate generated by specific 3-O-sulfotransferases. The viral ligand for these receptors is glycoprotein D (gD). To def...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2004-12, Vol.101 (49), p.17252-17257
Main Authors: Yoon, Miri, Spear, Patricia G.
Format: Article
Language:English
Subjects:
Animals
Binding Sites
Biological Sciences
Cell lines
Chlorocebus aethiops
Genetic mutation
Glycoproteins
Heparin - analogs & derivatives
Heparin - chemistry
Herpes simplex virus 1
Herpes viruses
Herpesvirus
Herpesvirus 1, Human - genetics
Herpesvirus 1, Human - pathogenicity
Human herpesvirus 1
Libraries
Microbiology
Mutagenesis
Mutation
Mutation, Missense
Open reading frames
Peptide Library
Plasmids
Protein Binding
Protein Conformation
Proteoglycans - chemistry
Receptors
Receptors, Tumor Necrosis Factor - chemistry
Receptors, Tumor Necrosis Factor, Member 14
Receptors, Virus - chemistry
Vero Cells
Viral Envelope Proteins - chemistry
Viral Envelope Proteins - genetics
Viral Envelope Proteins - physiology
Viruses
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Description
Summary:Herpes simplex virus type 1 (HSV-1) can enter cells expressing any one of multiple entry receptors, including the herpesvirus entry mediator (HVEM), nectin-1, and sites in heparan sulfate generated by specific 3-O-sulfotransferases. The viral ligand for these receptors is glycoprotein D (gD). To define structural requirements for functional interactions of gD with its receptors and to obtain viral mutants altered for receptor usage, we generated a library of HSV-1 mutants with random mutations in the gD gene. Viral isolates selected on a monkey cell line (Vero) were screened for the loss of ability to infect cells expressing each of the HSV-1 receptors. The 10 HSV-1 mutants obtained had 12 mutations in gD, affecting 11 amino acids. All mutations reduced or abrogated viral entry through HVEM and 3-O-sulfated heparan sulfate, indicating that similar features of gD are critical for functional interactions with both these receptors. None of the mutations reduced viral entry through nectin-1, whereas a subset of the mutations conferred ability to use nectin-2 as an entry receptor. These and other results show that features of gD, including conformation of the N terminus, critical for functional interactions with HVEM/3-O-sulfated heparan sulfate, differ from those critical for interactions with nectin-1.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.0407892101