{
  "abstract": "Herpes simplex virus 1 (HSV‑1) establishes infection through coordinated interactions of multiple viral glycoproteins with host cell receptors. Interactions between the viral glycoprotein D (gD) and the immunomodulatory cellular receptor HVEM (herpesvirus entry mediator) critically influence HSV‑1 latency and reactivation by regulating both receptor engagement and immunomodulatory signaling in the corneal epithelium, immune cells, and neurons. The underlying mechanisms of these phenotypes remain poorly understood because no tools selectively block their interaction. To dissect the contributions of individual amino acids (aa) to HVEM binding, we co‑immunoprecipitated wild‑type (WT) and mutant (Q27P and Q27A‑L28A‑T29A) forms of gD that disrupt gD binding to HVEM. We found that gD aa Q27-T29 were critical for HVEM binding, with the triple mutant showing greater disruption of HVEM binding than the Q27P single mutation, as assessed by co-immunoprecipitation and immunostaining. To map gD binding to HVEM and assess functional relevance, we constructed two recombinant viruses: a single-amino-acid (aa) gD mutation (v27 virus) and a triple-aa mutation in gD (v27-29 virus). Replication of both mutant viruses was comparable to that of the WT virus in four mammalian cell lines, except in Neuro2a cells. However, using confocal microscopy, HVEM protein expression was reduced in v27-29-infected Neuro2a cells, with no gD colocalization observed. Although Neuro2a cells infected with the single gD v27 mutant also showed less HVEM-gD colocalization than Neuro2a cells infected with WT virus, gD protein levels were not affected. RT‑PCR analysis of Neuro2a cells infected with single and triple gD mutants showed significantly less expression of viral glycoproteins (gB, gC, gD), host receptors (HVEM, nectin‑1, 3‑O‑sulfated heparan sulfate), and HVEM ligands (BTLA, LIGHT, Ltα, CD160), with the most pronounced effects observed for the triple v27‑29 mutant virus. Together, these results show that aa Q27-T29, rather than aa Q27 alone, are essential to completely block gD binding to HVEM and, consequently, affect overall receptor homeostasis. Author summary: Previously, we demonstrated that loss of HVEM impairs HSV-1 reactivation without significantly affecting primary viral replication. As a member of the TNF receptor superfamily, HVEM functions as a key immunomodulatory receptor and a critical entry mediator for HSV-1 through its interaction with glycoprotein D (gD). Beyond facilitating viral entry, gD–HVEM engagement can influence host immune signaling pathways, thereby shaping the local immune environment during infection. In this study, we identify amino acids 27–29 of gD as essential determinants for HVEM binding. These findings provide mechanistic insight into how HSV-1 exploits specific receptor interactions not only to gain entry but also to modulate immune responses. Disruption of the gD–HVEM interface, therefore, represents a promising strategy to limit viral spread and reactivation while potentially preserving antigen presentation and immune recognition.",
  "abstract_shown": true,
  "affiliations": [
    "Center for Neurobiology and Vaccine Development, Ophthalmology Research, Department of Surgery, Cedars-Sinai Health Sciences University, Cedars-Sinai Medical Center, Los Angeles, California, United States of America"
  ],
  "article_type": "Research Article",
  "arxiv": null,
  "author_total": 5,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Deepak Arya"
    },
    {
      "name": "Kati Tormanen"
    },
    {
      "name": "Jay J. Oh"
    },
    {
      "name": "Shaohui Wang"
    },
    {
      "name": "Homayon Ghiasi"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/emwz7cxg8zt4z.bib",
  "cited_by": 0,
  "doi": "10.1371/journal.ppat.1014566",
  "fulltext_url": "https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1014566",
  "id": "emwz7cxg8zt4z",
  "issns": [
    "1553-7374"
  ],
  "landing_url": "https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1014566",
  "language": null,
  "licence": "CC-BY-4.0",
  "licence_label": "CC BY 4.0",
  "licence_source": "journal",
  "licence_url": "https://creativecommons.org/licenses/by/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "openalex",
      "2713|Herpesvirus Infections and Treatments"
    ],
    [
      "plos",
      "/Biology and life sciences/Biochemistry/Glycobiology/Glycoproteins/Virus glycoproteins"
    ],
    [
      "plos",
      "/Biology and life sciences/Cell biology/Cell physiology/Cell binding"
    ],
    [
      "plos",
      "/Biology and life sciences/Genetics/Gene expression/Viral gene expression"
    ],
    [
      "plos",
      "/Biology and life sciences/Genetics/Microbial genetics/Viral genetics/Viral gene expression"
    ],
    [
      "plos",
      "/Biology and life sciences/Genetics/Mutation/Substitution mutation"
    ],
    [
      "plos",
      "/Biology and life sciences/Microbiology/Microbial mutation"
    ],
    [
      "plos",
      "/Biology and life sciences/Microbiology/Virology/Viral genetics/Viral gene expression"
    ],
    [
      "plos",
      "/Biology and life sciences/Microbiology/Virology/Viral replication"
    ],
    [
      "plos",
      "/Biology and life sciences/Molecular biology/Molecular biology techniques/Transfection"
    ],
    [
      "plos",
      "/Research and analysis methods/Chemical characterization/Binding analysis/Cell binding assay"
    ],
    [
      "plos",
      "/Research and analysis methods/Molecular biology techniques/Transfection"
    ]
  ],
  "openalex": "W7211944824",
  "pdf_url": "https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1014566&type=printable",
  "pmcid": null,
  "pmid": null,
  "published": "2026-09-08",
  "publisher": "Public Library of Science",
  "ris": "https://research.draconitic.com/paper/emwz7cxg8zt4z.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:17:56Z",
      "native_id": "10.1371/journal.ppat.1014566",
      "source": "plos",
      "url": "https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1014566"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "plos:gene expression",
      "subfield": "molecular-biology"
    },
    {
      "field": "biology",
      "origin": "plos:genetics",
      "subfield": "genetics-genomics"
    },
    {
      "field": "biology",
      "origin": "plos:microbiology",
      "subfield": "microbiology"
    }
  ],
  "title": "Complete disruption of gD binding to HVEM alters virus replication and host receptor homeostasis",
  "title_text": "Complete disruption of gD binding to HVEM alters virus replication and host receptor homeostasis",
  "type": "peer-reviewed",
  "updated_at": "2026-10-30T12:17:56Z",
  "url": "https://research.draconitic.com/paper/emwz7cxg8zt4z",
  "venue": "PLOS Pathogens",
  "versions": []
}