ADAP2 Is an Interferon Stimulated Gene That Restricts RNA Virus Entry

• Published in: PLoS pathogens 2015-09, Vol.11 (9), p.e1005150-e1005150
• Main Authors: Shu, Qian, Lennemann, Nicholas J, Sarkar, Saumendra N, Sadovsky, Yoel, Coyne, Carolyn B
• Format: Article
• Language: English
• Subjects: ADP-Ribosylation Factors - agonists; ADP-Ribosylation Factors - genetics; ADP-Ribosylation Factors - metabolism; Animals; Cell Line; Cercopithecus aethiops; Colleges & universities; Dengue fever; Dengue Virus - immunology; Dengue Virus - physiology; Enterovirus B, Human - immunology; Enterovirus B, Human - physiology; Gene expression; Genetic aspects; GTPase-Activating Proteins - agonists; GTPase-Activating Proteins - genetics; GTPase-Activating Proteins - metabolism; Health aspects; Host-virus relationships; Humans; Infections; Interferon; Interferon-beta - metabolism; Mutation; Observations; Pathogenesis; Phosphatidylinositol Phosphates - metabolism; Pinocytosis; Prevention; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - metabolism; RNA viruses; Sendai virus - immunology; Sendai virus - physiology; STAT1 Transcription Factor - genetics; STAT1 Transcription Factor - metabolism; Vero Cells; Vesiculovirus - immunology; Vesiculovirus - physiology; Viral infections; Virion - immunology; Virion - physiology; Virus Internalization; Virus Replication; Viruses
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/journal.ppat.1005150

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.