title: Interferon-induced antiviral protein MxA interacts with the cellular RNA helicases UAP56 and URH49.
abstract: Mx proteins are a family of large GTPases that are induced exclusively by interferon-α/β and have a broad antiviral activity against several viruses, including influenza A virus (IAV). Although the antiviral activities of mouse Mx1 and human MxA have been studied extensively, the molecular mechanism of action remains largely unsolved. Because no direct interaction between Mx proteins and IAV proteins or RNA had been demonstrated so far, we addressed the question of whether Mx protein would interact with cellular proteins required for efficient replication of IAV. Immunoprecipitation of MxA revealed its association with two closely related RNA helicases, UAP56 and URH49. UAP56 and its paralog URH49 play an important role in IAV replication and are involved in nuclear export of IAV mRNAs and prevention of dsRNA accumulation in infected cells. In vitro binding assays with purified recombinant proteins revealed that MxA formed a direct complex with the RNA helicases. In addition, recombinant mouse Mx1 was also able to bind to UAP56 or URH49. Furthermore, the complex formation between cytoplasmic MxA and UAP56 or URH49 occurred in the perinuclear region, whereas nuclear Mx1 interacted with UAP56 or URH49 in distinct dots in the nucleus. Taken together, our data reveal that Mx proteins exerting antiviral activity can directly bind to the two cellular DExD/H box RNA helicases UAP56 and URH49. Moreover, the observed subcellular localization of the Mx-RNA helicase complexes coincides with the subcellular localization, where human MxA and mouse Mx1 proteins act antivirally. On the basis of these data, we propose that Mx proteins exert their antiviral activity against IAV by interfering with the function of the RNA helicases UAP56 and URH49.
authors.affiliation: Institute of Medical Virology, University of Zurich, 8057 Zürich, Switzerland.
authors.forename: Christian
authors.initials: C
authors.lastname: Wisskirchen
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Ludersdorfer
authors.forename: Dominik A
authors.initials: DA
authors.lastname: Müller
authors.forename: Eva
authors.initials: E
authors.lastname: Moritz
authors.forename: Jovan
authors.initials: J
authors.lastname: Pavlovic
mesh_terms: D016475:3T3 Cells; D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism* / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D053487:DEAD-box RNA Helicases / Q000378:metabolism*; D019084:Fluorescent Antibody Technique, Indirect; D019204:GTP-Binding Proteins / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D009980:Influenza A virus / Q000378:metabolism; D007372:Interferons / Q000378:metabolism*; D051379:Mice; D064547:Myxovirus Resistance Proteins; D011485:Protein Binding; D020365:RNA Helicases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000998:Antiviral Agents; D053487:DEAD-box RNA Helicases; D019204:GTP-Binding Proteins; D049452:Green Fluorescent Proteins; D007372:Interferons; D064547:Myxovirus Resistance Proteins; D020365:RNA Helicases; C579071:MX1 protein, human; C579070:Mx1 protein, mouse; C518885:Ddx39b protein, mouse; C568467:Ddx39a protein, mouse; C498428:DDX39A protein, human; C487886:DDX39B protein, human