title: Interactome analysis of herpes simplex virus 1 envelope glycoprotein H.
abstract: Herpes simplex virus 1 (HSV-1) envelope glycoprotein H (gH) is important for viral entry into cells and nuclear egress of nucleocapsids. To clarify additional novel roles of gH during HSV-1 replication, host cell proteins that interact with gH were screened for by tandem affinity purification coupled with mass spectrometry-based proteomics in 293T cells transiently expressing gH. This screen identified 123 host cell proteins as potential gH interactors. Of these proteins, general control nonderepressive-1 (GCN1), a trans-acting positive effector of GCN2 kinase that regulates phosphorylation of the α subunit of translation initiation factor 2 (eIF2α), was subsequently confirmed to interact with gH in HSV-1-infected cells. eIF2α phosphorylation is known to downregulate protein synthesis, and various viruses have evolved mechanisms to prevent the accumulation of phosphorylated eIF2α in infected cells. Here, it was shown that GCN1 knockdown reduces phosphorylation of eIF2α in HSV-1-infected cells and that the gH-null mutation increases eIF2α in HSV-1-infected cells, whereas gH overexpression in the absence of other HSV-1 proteins reduces eIF2α phosphorylation. These findings suggest that GCN1 can regulate eIF2α phosphorylation in HSV-1-infected cells and that the GCN1-binding viral partner gH is necessary and sufficient to prevent the accumulation of phosphorylated eIF2α. Our database of 123 host cell proteins potentially interacting with gH will be useful for future studies aimed at unveiling further novel functions of gH and the roles of cellular proteins in HSV-1-infected cells.
authors: Division of Molecular Virology, Department of Microbiology and Immunology.|Department of Infectious Disease Control, International Research Center for Infectious Diseases.
authors: Yoshitaka
authors: Y
authors: Hirohata
authors: Division of Molecular Virology, Department of Microbiology and Immunology.|Department of Infectious Disease Control, International Research Center for Infectious Diseases.
authors: Akihisa
authors: A
authors: Kato
authors: Medical Proteomics Laboratory, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan.
authors: Masaaki
authors: M
authors: Oyama
authors: Medical Proteomics Laboratory, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan.
authors: Hiroko
authors: H
authors: Kozuka-Hata
authors: Division of Molecular Virology, Department of Microbiology and Immunology.|Department of Infectious Disease Control, International Research Center for Infectious Diseases.
authors: Naoto
authors: N
authors: Koyanagi
authors: Division of Molecular Virology, Department of Microbiology and Immunology.|Department of Infectious Disease Control, International Research Center for Infectious Diseases.
authors: Jun
authors: J
authors: Arii
authors: Division of Molecular Virology, Department of Microbiology and Immunology.|Department of Infectious Disease Control, International Research Center for Infectious Diseases.
authors: Yasushi
authors: Y
authors: Kawaguchi
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002460:Cell Line; D002846:Chromatography, Affinity; D004847:Epithelial Cells / Q000737:chemistry / Q000821:virology; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000502:physiology*; D006801:Humans; D013058:Mass Spectrometry; D010766:Phosphorylation; D011485:Protein Binding; D060066:Protein Interaction Maps; D011499:Protein Processing, Post-Translational; D016601:RNA-Binding Proteins; D015534:Trans-Activators; D014759:Viral Envelope Proteins / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D015852:Eukaryotic Initiation Factor-2; C000605933:GCN1 protein, human; D016601:RNA-Binding Proteins; D015534:Trans-Activators; D014759:Viral Envelope Proteins; C044200:glycoprotein H, herpes simplex virus type 1
keywords: animal DNA virus; glycoprotein H; herpes simplex virus