title: An Interferon Regulated MicroRNA Provides Broad Cell-Intrinsic Antiviral Immunity through Multihit Host-Directed Targeting of the Sterol Pathway.
abstract: In invertebrates, small interfering RNAs are at the vanguard of cell-autonomous antiviral immunity. In contrast, antiviral mechanisms initiated by interferon (IFN) signaling predominate in mammals. Whilst mammalian IFN-induced miRNA are known to inhibit specific viruses, it is not known whether host-directed microRNAs, downstream of IFN-signaling, have a role in mediating broad antiviral resistance. By performing an integrative, systematic, global analysis of RNA turnover utilizing 4-thiouridine labeling of newly transcribed RNA and pri/pre-miRNA in IFN-activated macrophages, we identify a new post-transcriptional viral defense mechanism mediated by miR-342-5p. On the basis of ChIP and site-directed promoter mutagenesis experiments, we find the synthesis of miR-342-5p is coupled to the antiviral IFN response via the IFN-induced transcription factor, IRF1. Strikingly, we find miR-342-5p targets mevalonate-sterol biosynthesis using a multihit mechanism suppressing the pathway at different functional levels: transcriptionally via SREBF2, post-transcriptionally via miR-33, and enzymatically via IDI1 and SC4MOL. Mass spectrometry-based lipidomics and enzymatic assays demonstrate the targeting mechanisms reduce intermediate sterol pathway metabolites and total cholesterol in macrophages. These results reveal a previously unrecognized mechanism by which IFN regulates the sterol pathway. The sterol pathway is known to be an integral part of the macrophage IFN antiviral response, and we show that miR-342-5p exerts broad antiviral effects against multiple, unrelated pathogenic viruses such Cytomegalovirus and Influenza A (H1N1). Metabolic rescue experiments confirm the specificity of these effects and demonstrate that unrelated viruses have differential mevalonate and sterol pathway requirements for their replication. This study, therefore, advances the general concept of broad antiviral defense through multihit targeting of a single host pathway.
authors.affiliation: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom.|SynthSys at Edinburgh University, The King's Buildings, Edinburgh, United Kingdom.
authors.forename: Kevin A
authors.initials: KA
authors.lastname: Robertson
authors.affiliation: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Wei Yuan
authors.initials: WY
authors.lastname: Hsieh
authors.affiliation: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom.|SynthSys at Edinburgh University, The King's Buildings, Edinburgh, United Kingdom.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Forster
authors.affiliation: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom.|École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Blanc
authors.affiliation: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Hongjin
authors.initials: H
authors.lastname: Lu
authors.affiliation: Institute of Mass Spectrometry, College of Medicine, Grove Building, Swansea University, Singleton Park, Swansea, United Kingdom.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Crick
authors.affiliation: Institute of Mass Spectrometry, College of Medicine, Grove Building, Swansea University, Singleton Park, Swansea, United Kingdom.
authors.forename: Eylan
authors.initials: E
authors.lastname: Yutuc
authors.affiliation: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom.|Northern Ireland Centre for Stratified Medicine, University of Ulster, C-Tric, Altnagelvin Campus, Londonderry, Ireland.
authors.forename: Steven
authors.initials: S
authors.lastname: Watterson
authors.affiliation: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom.|Centre for Integrative Physiology, Edinburgh, United Kingdom.
authors.forename: Kimberly
authors.initials: K
authors.lastname: Martin
authors.affiliation: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Samantha J
authors.initials: SJ
authors.lastname: Griffiths
authors.affiliation: EMBL - European Bioinformatics Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, United Kingdom.
authors.forename: Anton J
authors.initials: AJ
authors.lastname: Enright
authors.affiliation: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Mami
authors.initials: M
authors.lastname: Yamamoto
authors.affiliation: The Institute of Genetics and Molecular Medicine, Western General Hospital, Edinburgh, United Kingdom.
authors.forename: Madapura M
authors.initials: MM
authors.lastname: Pradeepa
authors.affiliation: Integrated DNA Technologies, Coralville, Iowa, United States of America.
authors.forename: Kimberly A
authors.initials: KA
authors.lastname: Lennox
authors.affiliation: Integrated DNA Technologies, Coralville, Iowa, United States of America.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Behlke
authors.affiliation: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Simon
authors.initials: S
authors.lastname: Talbot
authors.affiliation: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Haas
authors.affiliation: Department of Medicine, University of Cambridge, Cambridge, United Kingdom.|Institute of Virology, University of Würzburg, Würzburg, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Dölken
authors.affiliation: Institute of Mass Spectrometry, College of Medicine, Grove Building, Swansea University, Singleton Park, Swansea, United Kingdom.
authors.forename: William J
authors.initials: WJ
authors.lastname: Griffiths
authors.affiliation: Institute of Mass Spectrometry, College of Medicine, Grove Building, Swansea University, Singleton Park, Swansea, United Kingdom.
authors.forename: Yuqin
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Institut d'Investigacions Biomèdiques August Pi i Sunyer, Barcelona, Spain.|Immunology Unit, Department of Cell Biology, Immunology, and Neurosciences, Medical School, University of Barcelona, Barcelona, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Angulo
authors.affiliation: Division of Pathway Medicine, University of Edinburgh, Edinburgh, United Kingdom.|SynthSys at Edinburgh University, The King's Buildings, Edinburgh, United Kingdom.
authors.forename: Peter
authors.initials: P
authors.lastname: Ghazal
mesh_terms: D000818:Animals; D050856:Interferon Regulatory Factor-1 / Q000378:metabolism*; D007372:Interferons / Q000502:physiology*; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000378:metabolism*; D013261:Sterols / Q000096:biosynthesis*; D014777:Virus Diseases / Q000276:immunology*
chemical_list: D050856:Interferon Regulatory Factor-1; C494258:Irf1 protein, mouse; D035683:MicroRNAs; C581863:Mirn342 microRNA, mouse; D013261:Sterols; D007372:Interferons