title: ATP hydrolysis by the viral RNA sensor RIG-I prevents unintentional recognition of self-RNA.
abstract: The cytosolic antiviral innate immune sensor RIG-I distinguishes 5' tri- or diphosphate containing viral double-stranded (ds) RNA from self-RNA by an incompletely understood mechanism that involves ATP hydrolysis by RIG-I's RNA translocase domain. Recently discovered mutations in ATPase motifs can lead to the multi-system disorder Singleton-Merten Syndrome (SMS) and increased interferon levels, suggesting misregulated signaling by RIG-I. Here we report that SMS mutations phenocopy a mutation that allows ATP binding but prevents hydrolysis. ATPase deficient RIG-I constitutively signals through endogenous RNA and co-purifies with self-RNA even from virus infected cells. Biochemical studies and cryo-electron microscopy identify a 60S ribosomal expansion segment as a dominant self-RNA that is stably bound by ATPase deficient RIG-I. ATP hydrolysis displaces wild-type RIG-I from this self-RNA but not from 5' triphosphate dsRNA. Our results indicate that ATP-hydrolysis prevents recognition of self-RNA and suggest that SMS mutations lead to unintentional signaling through prolonged RNA binding.
authors.affiliation: Gene Center, Department of Biochemistry, Ludwig Maximilian University of Munich, Munich, Germany.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Lässig
authors.affiliation: Gene Center, Department of Biochemistry, Ludwig Maximilian University of Munich, Munich, Germany.
authors.forename: Sarah
authors.initials: S
authors.lastname: Matheisl
authors.affiliation: Max von Pettenkofer-Institute, Gene Center, Ludwig Maximilian University of Munich, Munich, Germany.
authors.forename: Konstantin M J
authors.initials: KM
authors.lastname: Sparrer
authors.affiliation: Gene Center, Department of Biochemistry, Ludwig Maximilian University of Munich, Munich, Germany.
authors.forename: Carina C
authors.initials: CC
authors.lastname: de Oliveira Mann
authors.affiliation: Gene Center, Department of Biochemistry, Ludwig Maximilian University of Munich, Munich, Germany.
authors.forename: Manuela
authors.initials: M
authors.lastname: Moldt
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, United States.|Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, United States.
authors.forename: Jenish R
authors.initials: JR
authors.lastname: Patel
authors.affiliation: Institute for Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, University of Bonn, Bonn, Germany.
authors.forename: Marion
authors.initials: M
authors.lastname: Goldeck
authors.affiliation: Institute for Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, University of Bonn, Bonn, Germany.
authors.forename: Gunther
authors.initials: G
authors.lastname: Hartmann
authors.affiliation: Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, United States.|Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, United States.|Department of Medicine, Division of Infectious Diseases, Icahn School of Medicine at Mount Sinai, New York, United States.
authors.forename: Adolfo
authors.initials: A
authors.lastname: García-Sastre
authors.affiliation: Institute of Molecular Medicine, University Hospital Bonn, University of Bonn, Bonn, Germany.
authors.forename: Veit
authors.initials: V
authors.lastname: Hornung
authors.affiliation: Max von Pettenkofer-Institute, Gene Center, Ludwig Maximilian University of Munich, Munich, Germany.
authors.forename: Karl-Klaus
authors.initials: KK
authors.lastname: Conzelmann
authors.affiliation: Gene Center, Department of Biochemistry, Ludwig Maximilian University of Munich, Munich, Germany.|Center for Integrated Protein Science Munich, Munich, Germany.
authors.forename: Roland
authors.initials: R
authors.lastname: Beckmann
authors.affiliation: Gene Center, Department of Biochemistry, Ludwig Maximilian University of Munich, Munich, Germany.|Center for Integrated Protein Science Munich, Munich, Germany.
authors.forename: Karl-Peter
authors.identifier: http://orcid.org/0000-0002-4528-8357
authors.initials: KP
authors.lastname: Hopfner
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D002460:Cell Line; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D012367:RNA, Viral / Q000378:metabolism*; D011971:Receptors, Immunologic; D013379:Substrate Specificity
chemical_list: D012367:RNA, Viral; D011971:Receptors, Immunologic; D000255:Adenosine Triphosphate; C452820:RIGI protein, human; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases
keywords: ATPase domain; RIG-I; RLR; Singleton-Merten Syndrome; autoimmune response / disease; cell biology; human; immunology; innate immune system