title: ADAP2 Is an Interferon Stimulated Gene That Restricts RNA Virus Entry.
abstract: Interferon stimulated genes (ISGs) target viruses at various stages of their infectious life cycles, including at the earliest stage of viral entry. Here we identify ArfGAP with dual pleckstrin homology (PH) domains 2 (ADAP2) as a gene upregulated by type I IFN treatment in a STAT1-dependent manner. ADAP2 functions as a GTPase-activating protein (GAP) for Arf6 and binds to phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P3) and PI(3,4)P2. We show that overexpression of ADAP2 suppresses dengue virus (DENV) and vesicular stomatitis virus (VSV) infection in an Arf6 GAP activity-dependent manner, while exerting no effect on coxsackievirus B (CVB) or Sendai virus (SeV) replication. We further show that ADAP2 expression induces macropinocytosis and that ADAP2 strongly associates with actin-enriched membrane ruffles and with Rab8a- and LAMP1-, but not EEA1- or Rab7-, positive vesicles. Utilizing two techniques--light-sensitive neutral red (NR)-containing DENV and fluorescence assays for virus internalization--we show that ADAP2 primarily restricts DENV infection at the stage of virion entry and/or intracellular trafficking and that incoming DENV and VSV particles associate with ADAP2 during their entry. Taken together, this study identifies ADAP2 as an ISG that exerts antiviral effects against RNA viruses by altering Arf6-mediated trafficking to disrupt viral entry.
authors.affiliation: Tsinghua University School of Medicine, Beijing, China; Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Qian
authors.initials: Q
authors.lastname: Shu
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Lennemann
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America; University of Pittsburgh Cancer Institute, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Saumendra N
authors.initials: SN
authors.lastname: Sarkar
authors.affiliation: Magee-Womens Research Institute, Department of Obstetrics, Gynecology, and Reproductive Science, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Yoel
authors.initials: Y
authors.lastname: Sadovsky
authors.affiliation: Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America; Magee-Womens Research Institute, Department of Obstetrics, Gynecology, and Reproductive Science, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Carolyn B
authors.initials: CB
authors.lastname: Coyne
mesh_terms: D000089982:ADP-Ribosylation Factor 6; D020727:ADP-Ribosylation Factors / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D003716:Dengue Virus / Q000276:immunology / Q000502:physiology*; D029822:Enterovirus B, Human / Q000276:immunology / Q000502:physiology; D020690:GTPase-Activating Proteins / Q000819:agonists* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D016899:Interferon-beta / Q000378:metabolism*; D009154:Mutation; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D010873:Pinocytosis; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D050794:STAT1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D029082:Sendai virus / Q000276:immunology / Q000502:physiology; D014709:Vero Cells; D018116:Vesiculovirus / Q000276:immunology / Q000502:physiology*; D014771:Virion / Q000276:immunology / Q000502:physiology; D053586:Virus Internalization; D014779:Virus Replication
chemical_list: C470610:ADAP2 protein, human; D000089982:ADP-Ribosylation Factor 6; D020690:GTPase-Activating Proteins; D018129:Phosphatidylinositol Phosphates; D011993:Recombinant Fusion Proteins; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; C060974:phosphatidylinositol 3,4,5-triphosphate; C060980:phosphatidylinositol 3,4-diphosphate; D016899:Interferon-beta; D020727:ADP-Ribosylation Factors; C000715348:ARF6 protein, human