title: In vivo ligands of MDA5 and RIG-I in measles virus-infected cells.
abstract: RIG-I-like receptors (RLRs: RIG-I, MDA5 and LGP2) play a major role in the innate immune response against viral infections and detect patterns on viral RNA molecules that are typically absent from host RNA. Upon RNA binding, RLRs trigger a complex downstream signaling cascade resulting in the expression of type I interferons and proinflammatory cytokines. In the past decade extensive efforts were made to elucidate the nature of putative RLR ligands. In vitro and transfection studies identified 5'-triphosphate containing blunt-ended double-strand RNAs as potent RIG-I inducers and these findings were confirmed by next-generation sequencing of RIG-I associated RNAs from virus-infected cells. The nature of RNA ligands of MDA5 is less clear. Several studies suggest that double-stranded RNAs are the preferred agonists for the protein. However, the exact nature of physiological MDA5 ligands from virus-infected cells needs to be elucidated. In this work, we combine a crosslinking technique with next-generation sequencing in order to shed light on MDA5-associated RNAs from human cells infected with measles virus. Our findings suggest that RIG-I and MDA5 associate with AU-rich RNA species originating from the mRNA of the measles virus L gene. Corresponding sequences are poorer activators of ATP-hydrolysis by MDA5 in vitro, suggesting that they result in more stable MDA5 filaments. These data provide a possible model of how AU-rich sequences could activate type I interferon signaling.
authors.affiliation: Gene Center and Department of Biochemistry, Ludwig-Maximilians University Munich, Munich, Germany.
authors.forename: Simon
authors.initials: S
authors.lastname: Runge
authors.affiliation: Max von Pettenkofer-Institute, Gene Center, Ludwig-Maximilians University Munich, Munich, Germany.
authors.forename: Konstantin M J
authors.initials: KM
authors.lastname: Sparrer
authors.affiliation: Gene Center and Department of Biochemistry, Ludwig-Maximilians University Munich, Munich, Germany.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Lässig
authors.affiliation: Gene Center and Department of Biochemistry, Ludwig-Maximilians University Munich, Munich, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Hembach
authors.affiliation: Center for the Study of Hepatitis C, Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, New York, United States of America.
authors.forename: Alina
authors.initials: A
authors.lastname: Baum
authors.affiliation: Department of Microbiology, Department of Medicine, Division of Infectious Diseases and Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, New York, United States of America.
authors.forename: Adolfo
authors.initials: A
authors.lastname: García-Sastre
authors.affiliation: Gene Center and Department of Biochemistry, Ludwig-Maximilians University Munich, Munich, Germany; Center for Integrated Protein Science Munich, Munich, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Söding
authors.affiliation: Max von Pettenkofer-Institute, Gene Center, Ludwig-Maximilians University Munich, Munich, Germany.
authors.forename: Karl-Klaus
authors.initials: KK
authors.lastname: Conzelmann
authors.affiliation: Gene Center and Department of Biochemistry, Ludwig-Maximilians University Munich, Munich, Germany; Center for Integrated Protein Science Munich, Munich, Germany.
authors.forename: Karl-Peter
authors.initials: KP
authors.lastname: Hopfner
mesh_terms: D045744:Cell Line, Tumor; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D000072640:Interferon-Induced Helicase, IFIH1; D008457:Measles / Q000235:genetics / Q000378:metabolism*; D008459:Measles virus / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism*; D011971:Receptors, Immunologic; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D012333:RNA, Messenger; D012367:RNA, Viral; D011971:Receptors, Immunologic; D014764:Viral Proteins; C452820:RIGI protein, human; C452815:IFIH1 protein, human; D000071457:DEAD Box Protein 58; D053487:DEAD-box RNA Helicases; D000072640:Interferon-Induced Helicase, IFIH1
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