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RIG-I-Like Receptors: Their Regulation and Roles in RNA Sensing

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RIG-I-Like Receptors: Their Regulation and Roles in RNA Sensing REVIEWS RIG- I- like receptors: their regulation and roles in RNA sensing Jan Rehwinkel 1 ✉ and Michaela U. Gack 2 ✉ Abstract | Retinoic acid- inducible gene I (RIG- I)- like receptors (RLRs) are key sensors of virus infection, mediating the transcriptional induction of type I interferons and other genes that collectively establish an antiviral host response. Recent studies have revealed that both viral and host- derived RNAs can trigger RLR activation; this can lead to an effective antiviral response but also immunopathology if RLR activities are uncontrolled. In this Review , we discuss recent advances in our understanding of the types of RNA sensed by RLRs in the contexts of viral infection, malignancies and autoimmune diseases. We further describe how the activity of RLRs is controlled by host regulatory mechanisms, including RLR-interacting proteins, post- translational modifications and non- coding RNAs. Finally , we discuss key outstanding questions in the RLR field, including how our knowledge of RLR biology could be translated into new therapeutics. Toll- like receptors Type I interferons are highly potent cytokines that were Their expression is rapidly induced by several innate (TLRs). A family of initially identified for their essential role in antiviral immune signalling pathways. These signalling events are membrane- bound innate defence1,2. They shape both innate and adaptive immune typically initiated by proteins called nucleic acid sensors immune receptors that responses and induce the expression of restriction fac- that monitor cells for unusual nucleic acids. For example, recognize various bacterial or virus- derived pathogen- tors, which are proteins that directly interfere with a step it is an indispensable step in the life cycle of any virus 3 associated molecular patterns. in the life cycle of a virus . Both beneficial and detrimen- to introduce a genome consisting of DNA or RNA into These sensors are crucial for tal roles for type I interferons have been described in the the infected cell. Nucleic acid sensors survey different eliciting an early innate context of bacterial infections and cancer2,4–6, and these subcellular compartments and include the endosomal immune response and for cytokines are also linked to the development of several Toll- like receptors (TLRs) TLR3, TLR7, TLR8 and TLR9; modulating adaptive immunity. autoinflammatory and autoimmune diseases7 and met- the cytosolic DNA sensor cyclic GMP–AMP synthase abolic syndromes8. Therefore, the study of the biology (cGAS); and retinoic acid- inducible gene I (RIG- I)- like of type I interferons has many implications for human receptors (RLRs)12,13. Once activated, these and other health and disease. sensors engage converging signalling cascades that The human genome encodes 17 different type I lead to transcriptional induction of the genes encoding interferons, including 13 types of interferon- α (IFNα), type I interferons and other immune genes (FIG. 1). as well as IFNβ, IFNε, IFNκ and IFNω. Type I inter- RLRs are RNA sensors localized in the cytosol12,13. ferons are secreted and then signal in an autocrine and This protein family encompasses three members: paracrine manner by binding to the type I interferon RIG- I, melanoma differentiation- associated protein 5 receptor (interferon- α/β receptor (IFNAR)) that acti- (MDA5) and laboratory of genetics and physiology 2 1Medical Research Council vates the intracellular Janus kinase–signal transducer (LGP2). All RLRs have a central helicase domain and Human Immunology Unit, and activator of transcription (JAK–STAT) pathway, a so- called carboxy- terminal domain (CTD) (FIG. 2a). Medical Research Council particularly STAT1 and STAT2 (REFS9,10). Upon phospho- These two domains work together to detect immunos- Weatherall Institute of Molecular Medicine, Radcliffe rylation, STAT1 and STAT2 combine with interferon reg- timulatory RNAs. RIG- I and MDA5 additionally have Department of Medicine, ulatory factor 9 (IRF9) to form the interferon-stimulated two amino-terminal caspase activation and recruitment University of Oxford, gene factor 3 (ISGF3) transcriptional complex11. ISGF3 domains (CARDs), which mediate downstream signal Oxford, UK. binds to interferon- stimulated response elements transduction. LGP2 lacks the CARDs and is widely 2Department of Microbiology, in type I interferon- dependent gene promoters and believed to regulate RIG-I and MDA5 (BOx 1). It is now The University of Chicago, thereby drives the expression of hundreds of type I over 15 years since RIG-I and MDA5 were discovered in Chicago, IL, USA. interferon- stimulated genes (ISGs). The proteins overexpression studies to induce type I interferons14,15. ✉e- mail: jan.rehwinkel@ imm.ox.ac.uk; mgack@ encoded by ISGs mediate the effects of type I interferons; Soon after their description, knockout studies in mice uchicago.edu for example, some ISGs encode virus restriction factors. and mouse- derived cells established that RIG- I and https://doi.org/10.1038/ Under homeostatic conditions, type I interferons MDA5 are essential for antiviral defence and type I inter- s41577-020-0288-3 are expressed at very low and often undetectable levels. feron induction in virus infection models16,17. It is now NATURE REVIEWS | IMMUNOLOGY VOLUME 20 | SEPTEMBER 2020 | 537 REVIEWS then induce transcription of the genes encoding type I Viruses interferons and other antiviral or immunoregulatory 12,13 (FIG. 2b) IFNAR genes . In this Review, we focus on recent developments in Type I interferon our understanding of the mechanisms by which RLRs DNA RNA are activated. First, we discuss the types and properties JAK–STAT of RNA molecules that trigger RIG-I and MDA5 activa- tion. Interestingly, these proteins not only detect viral Nucleic acid sensors Cancer RNA but also cellular RNAs that are unusual, mislo- Transcription of ISGs calized or misprocessed. Such disturbances of cellular Transcription of RNA homeostasis can be an indirect molecular signature type I interferon genes Bacteria of infection or occur in sterile inflammatory patholo- gies. In the second part of the article, we highlight the Autoimmunity regulation of RLR activity by host mechanisms such as post- translational modifications (PTMs), interacting proteins and cellular non- coding RNAs. Tight control Fig. 1 | The type I interferon system. Nucleic acid sensors of the innate immune system recognize unusual DNA and RNA molecules, for example, viral genomes. This results in of RLRs is important both to prevent detrimental type I 7 the triggering of an intracellular signalling cascade that transcriptionally induces the interferon responses in the absence of infection and to genes encoding type I interferons, which are subsequently secreted. Type I interferons allow rapid and sensitive recognition of pathogens. act in an autocrine (not shown) and paracrine manner by binding to their receptor, interferon-α /β receptor (IFNAR), which activates the Janus kinases (JAKs). These in turn RLR recognition of immunostimulatory RNA activate the transcription factors signal transducer and activator of transcription 1 RLRs are expressed in most cell types and are primarily (STAT1) and STAT2, leading to expression of interferon- stimulated genes (ISGs). Several located in the cytosol, although recent studies showed ISGs have direct antiviral effects. Type I interferons and ISGs also play important roles in that RIG- I may also localize to the cell nucleus27,28. They bacterial infections and cancer; these effects can be both beneficial and detrimental, induce type I interferon responses upon recognition of depending on the setting. Finally , production of type I interferons and ISGs over extended periods of time can lead to autoinflammatory and autoimmune diseases. RNA molecules that are usually not present in mamma- lian cells, for example, the genomes of RNA viruses. In this section, we discuss the properties that mark RNA clear that infections with almost all major virus families molecules as immunostimulatory and the principles are recognized by RLRs (TABLe 1). explaining why RLRs are not activated by the abun- Much work has also been done to delineate the dant RNA content of the cytosol of healthy cells. The structural biology of RLR activation. The structure mechanisms by which RLRs distinguish between nor- of full- length RIG- I without RNA ligand showed that mal RNAs found in cells under homeostatic conditions RIG- I is in a signalling- repressed, closed conformation and immunostimulatory RNAs are important for our that is mediated by specific contacts of CARD2 with an understanding of how type I interferon responses are inserting domain located between the two RecA-like hel- initiated during virus infection and in sterile patho- icase domains of RIG- I18. When bound to RNA ligand, logies. Furthermore, fundamental insights into RLR the helicase domain of RIG- I tightly wraps around the activation have led to the development of synthetic RNA in a C- clamp-like fashion. ‘Pincer’ helical elements RLR agonists, which may hold therapeutic value for viral found in the helicase of RIG-I connect to the CTD and and malignant diseases, as well as vaccination (BOx 2). are believed to trigger RNA binding- dependent con- formational changes in RIG- I that expose the CARDs Activation of RIG- I by viral RNAs. Since the discovery for signalling. The crystal structure of 2CARD- deleted of RIG- I, much research has been done to unravel the RIG- I, or the isolated CTD, bound to RNA revealed molecular and structural requirements
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