ATP hydrolysis by the viral RNA sensor RIG-I prevents unintentional recognition of self-RNA
Charlotte Lässig,
Sarah Matheisl,
Konstantin MJ Sparrer,
Carina C de Oliveira Mann,
Manuela Moldt,
Jenish R Patel,
Marion Goldeck,
Gunther Hartmann,
Adolfo García-Sastre,
Veit Hornung,
Karl-Klaus Conzelmann,
Roland Beckmann,
Karl-Peter Hopfner
Affiliations
Charlotte Lässig
Gene Center, Department of Biochemistry, Ludwig Maximilian University of Munich, Munich, Germany
Sarah Matheisl
Gene Center, Department of Biochemistry, Ludwig Maximilian University of Munich, Munich, Germany
Konstantin MJ Sparrer
Max von Pettenkofer-Institute, Gene Center, Ludwig Maximilian University of Munich, Munich, Germany
Carina C de Oliveira Mann
Gene Center, Department of Biochemistry, Ludwig Maximilian University of Munich, Munich, Germany
Manuela Moldt
Gene Center, Department of Biochemistry, Ludwig Maximilian University of Munich, Munich, Germany
Jenish R Patel
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
Marion Goldeck
Institute for Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, University of Bonn, Bonn, Germany
Gunther Hartmann
Institute for Clinical Chemistry and Clinical Pharmacology, University Hospital Bonn, University of Bonn, Bonn, Germany
Adolfo García-Sastre
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
Veit Hornung
Institute of Molecular Medicine, University Hospital Bonn, University of Bonn, Bonn, Germany
Karl-Klaus Conzelmann
Max von Pettenkofer-Institute, Gene Center, Ludwig Maximilian University of Munich, Munich, Germany
Roland Beckmann
Gene Center, Department of Biochemistry, Ludwig Maximilian University of Munich, Munich, Germany; Center for Integrated Protein Science Munich, Munich, Germany
Gene Center, Department of Biochemistry, Ludwig Maximilian University of Munich, Munich, Germany; Center for Integrated Protein Science Munich, Munich, Germany
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.
