title: Role of nucleotide binding and GTPase domain dimerization in dynamin-like myxovirus resistance protein A for GTPase activation and antiviral activity.
abstract: Myxovirus resistance (Mx) GTPases are induced by interferon and inhibit multiple viruses, including influenza and human immunodeficiency viruses. They have the characteristic domain architecture of dynamin-related proteins with an N-terminal GTPase (G) domain, a bundle signaling element, and a C-terminal stalk responsible for self-assembly and effector functions. Human MxA (also called MX1) is expressed in the cytoplasm and is partly associated with membranes of the smooth endoplasmic reticulum. It shows a protein concentration-dependent increase in GTPase activity, indicating regulation of GTP hydrolysis via G domain dimerization. Here, we characterized a panel of G domain mutants in MxA to clarify the role of GTP binding and the importance of the G domain interface for the catalytic and antiviral function of MxA. Residues in the catalytic center of MxA and the nucleotide itself were essential for G domain dimerization and catalytic activation. In pulldown experiments, MxA recognized Thogoto virus nucleocapsid proteins independently of nucleotide binding. However, both nucleotide binding and hydrolysis were required for the antiviral activity against Thogoto, influenza, and La Crosse viruses. We further demonstrate that GTP binding facilitates formation of stable MxA assemblies associated with endoplasmic reticulum membranes, whereas nucleotide hydrolysis promotes dynamic redistribution of MxA from cellular membranes to viral targets. Our study highlights the role of nucleotide binding and hydrolysis for the intracellular dynamics of MxA during its antiviral action.
authors.affiliation: From the Max-Delbrück Centrum für Molekulare Medizin, Robert-Rössle-Strasse 10, 13125 Berlin, Germany, the Institute of Chemistry and Biochemistry, Freie Universität Berlin, Takustrasse 6, 14195 Berlin, Germany.
authors.forename: Alexej
authors.initials: A
authors.lastname: Dick
authors.affiliation: the Institute of Virology, University Medical Center, Hermann-Herder-Strasse 11, 79104 Freiburg, Germany, the Spemann Graduate School of Biology and Medicine, University of Freiburg, Albertstrasse 19a, 79104 Freiburg, Germany, and.
authors.forename: Laura
authors.initials: L
authors.lastname: Graf
authors.affiliation: From the Max-Delbrück Centrum für Molekulare Medizin, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Olal
authors.affiliation: the Institute of Virology, University Medical Center, Hermann-Herder-Strasse 11, 79104 Freiburg, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: von der Malsburg
authors.affiliation: the Sun Yat-sen University Cancer Center, State Key Laboratory of Oncology in South China, and Collaborative Innovation Center for Cancer Medicine, Guangzhou 510060, China.
authors.forename: Song
authors.initials: S
authors.lastname: Gao
authors.affiliation: the Institute of Virology, University Medical Center, Hermann-Herder-Strasse 11, 79104 Freiburg, Germany, the Spemann Graduate School of Biology and Medicine, University of Freiburg, Albertstrasse 19a, 79104 Freiburg, Germany, and georg.kochs@uniklinik-freiburg.de.
authors.forename: Georg
authors.initials: G
authors.lastname: Kochs
authors.affiliation: From the Max-Delbrück Centrum für Molekulare Medizin, Robert-Rössle-Strasse 10, 13125 Berlin, Germany, the Institute of Chemistry and Biochemistry, Freie Universität Berlin, Takustrasse 6, 14195 Berlin, Germany, oliver.daumke@mdc-berlin.de.
authors.forename: Oliver
authors.initials: O
authors.lastname: Daumke
mesh_terms: D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation / Q000235:genetics; D020558:GTP Phosphohydrolases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D007425:Intracellular Membranes / Q000378:metabolism; D018062:La Crosse virus / Q000235:genetics / Q000378:metabolism; D064547:Myxovirus Resistance Proteins / Q000235:genetics / Q000378:metabolism*; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism*; D009975:Orthomyxoviridae / Q000235:genetics / Q000378:metabolism; D055503:Protein Multimerization / Q000502:physiology*; D017434:Protein Structure, Tertiary; D018117:Thogotovirus / Q000235:genetics / Q000378:metabolism*
chemical_list: C579071:MX1 protein, human; D064547:Myxovirus Resistance Proteins; D019590:Nucleocapsid Proteins; D020558:GTP Phosphohydrolases
keywords: G protein; Mx proteins; antiviral response; catalytic mechanism; dynamin-like protein; innate immunity; interferon; membrane; viral replication
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