An Interferon Regulated MicroRNA Provides Broad Cell-Intrinsic Antiviral Immunity through Multihit Host-Directed Targeting of the Sterol Pathway.

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...

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Main Authors: Kevin A Robertson, Wei Yuan Hsieh, Thorsten Forster, Mathieu Blanc, Hongjin Lu, Peter J Crick, Eylan Yutuc, Steven Watterson, Kimberly Martin, Samantha J Griffiths, Anton J Enright, Mami Yamamoto, Madapura M Pradeepa, Kimberly A Lennox, Mark A Behlke, Simon Talbot, Jürgen Haas, Lars Dölken, William J Griffiths, Yuqin Wang, Ana Angulo, Peter Ghazal
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2016-03-01
Series:PLoS Biology
Online Access:https://doi.org/10.1371/journal.pbio.1002364
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author Kevin A Robertson
Wei Yuan Hsieh
Thorsten Forster
Mathieu Blanc
Hongjin Lu
Peter J Crick
Eylan Yutuc
Steven Watterson
Kimberly Martin
Samantha J Griffiths
Anton J Enright
Mami Yamamoto
Madapura M Pradeepa
Kimberly A Lennox
Mark A Behlke
Simon Talbot
Jürgen Haas
Lars Dölken
William J Griffiths
Yuqin Wang
Ana Angulo
Peter Ghazal
author_facet Kevin A Robertson
Wei Yuan Hsieh
Thorsten Forster
Mathieu Blanc
Hongjin Lu
Peter J Crick
Eylan Yutuc
Steven Watterson
Kimberly Martin
Samantha J Griffiths
Anton J Enright
Mami Yamamoto
Madapura M Pradeepa
Kimberly A Lennox
Mark A Behlke
Simon Talbot
Jürgen Haas
Lars Dölken
William J Griffiths
Yuqin Wang
Ana Angulo
Peter Ghazal
author_sort Kevin A Robertson
collection DOAJ
description 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.
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spelling doaj-art-2a7e4b86c96d49edbfa01de93c1379492025-08-20T03:10:02ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852016-03-01143e100236410.1371/journal.pbio.1002364An Interferon Regulated MicroRNA Provides Broad Cell-Intrinsic Antiviral Immunity through Multihit Host-Directed Targeting of the Sterol Pathway.Kevin A RobertsonWei Yuan HsiehThorsten ForsterMathieu BlancHongjin LuPeter J CrickEylan YutucSteven WattersonKimberly MartinSamantha J GriffithsAnton J EnrightMami YamamotoMadapura M PradeepaKimberly A LennoxMark A BehlkeSimon TalbotJürgen HaasLars DölkenWilliam J GriffithsYuqin WangAna AnguloPeter GhazalIn 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.https://doi.org/10.1371/journal.pbio.1002364
spellingShingle Kevin A Robertson
Wei Yuan Hsieh
Thorsten Forster
Mathieu Blanc
Hongjin Lu
Peter J Crick
Eylan Yutuc
Steven Watterson
Kimberly Martin
Samantha J Griffiths
Anton J Enright
Mami Yamamoto
Madapura M Pradeepa
Kimberly A Lennox
Mark A Behlke
Simon Talbot
Jürgen Haas
Lars Dölken
William J Griffiths
Yuqin Wang
Ana Angulo
Peter Ghazal
An Interferon Regulated MicroRNA Provides Broad Cell-Intrinsic Antiviral Immunity through Multihit Host-Directed Targeting of the Sterol Pathway.
PLoS Biology
title An Interferon Regulated MicroRNA Provides Broad Cell-Intrinsic Antiviral Immunity through Multihit Host-Directed Targeting of the Sterol Pathway.
title_full An Interferon Regulated MicroRNA Provides Broad Cell-Intrinsic Antiviral Immunity through Multihit Host-Directed Targeting of the Sterol Pathway.
title_fullStr An Interferon Regulated MicroRNA Provides Broad Cell-Intrinsic Antiviral Immunity through Multihit Host-Directed Targeting of the Sterol Pathway.
title_full_unstemmed An Interferon Regulated MicroRNA Provides Broad Cell-Intrinsic Antiviral Immunity through Multihit Host-Directed Targeting of the Sterol Pathway.
title_short An Interferon Regulated MicroRNA Provides Broad Cell-Intrinsic Antiviral Immunity through Multihit Host-Directed Targeting of the Sterol Pathway.
title_sort interferon regulated microrna provides broad cell intrinsic antiviral immunity through multihit host directed targeting of the sterol pathway
url https://doi.org/10.1371/journal.pbio.1002364
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