Structure of STING bound to cyclic di-GMP reveals the mechanism of cyclic dinucleotide recognition by the immune system

Shu, Chen; Yi, Guanghui; Watts, Tylan; Kao, C. Cheng; Li, Pingwei

doi:10.1038/nsmb.2331

Cited by 253 publications

(239 citation statements)

References 23 publications

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“…Rather than converging on the activation of IRF3, as we had hypothesized, Ca 2ϩ signaling associated with membrane perturbation appears to be necessary for activation of STING. While no role for Ca 2ϩ in STING activation had been observed previously, the crystal structures of STING show an important Ca 2ϩ -binding pocket at the interface of 2 STING dimers (52,53). While these results suggest that Ca 2ϩ signaling is similarly upstream of MAVS activation following stimulation with enveloped RNA virus particles, we were unable to reproducibly detect MAVS activation in primary fibroblasts to test this hypothesis.…”

Section: Discussioncontrasting

confidence: 50%

Membrane Perturbation-Associated Ca ²⁺ Signaling and Incoming Genome Sensing Are Required for the Host Response to Low-Level Enveloped Virus Particle Entry

Hare

Collins

Mukherjee

et al. 2016

J Virol

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The type I interferon (IFN) response is an important aspect of innate antiviral defense, and the transcription factor IRF3 plays an important role in its induction. Membrane perturbation during fusion, a necessary step for enveloped virus particle entry, appears sufficient to induce transcription of a subset of IFN-stimulated genes (ISGs) in an IRF3-dependent, IFN-independent fashion. IRF3 is emerging as a central node in host cell stress responses, although it remains unclear how different forms of stress activate IRF3. Here, we investigated the minimum number of Sendai virus (SeV) and human cytomegalovirus (HCMV) particles required to activate IRF3 and trigger an antiviral response. We found that Ca 2؉ signaling associated with membrane perturbation and recognition of incoming viral genomes by cytosolic nucleic acid receptors are required to activate IRF3 in response to fewer than 13 particles of SeV and 84 particles of HCMV per cell. Moreover, it appears that Ca 2؉ signaling is important for activation of STING and IRF3 following HCMV particle entry, suggesting that Ca 2؉ signaling sensitizes cells to recognize genomes within incoming virus particles. To our knowledge, this is the first evidence that cytosolic nucleic acid sensors recognize genomes within incoming virus particles prior to virus replication. These studies highlight the exquisite sensitivity of the cellular response to low-level stimuli and suggest that virus particle entry is sensed as a stress signal. IMPORTANCEThe mechanism by which replicating viruses trigger IRF3 activation and type I IFN induction through the generation and accumulation of viral pathogen-associated molecular patterns has been well characterized. However, the mechanism by which enveloped virus particle entry mediates a stress response, leading to IRF3 activation and the IFN-independent response, remained elusive. Here, we find that Ca 2؉ signaling associated with membrane perturbation appears to sensitize cells to recognize genomes within incoming virus particles. To our knowledge, this is the first study to show that cytosolic receptors recognize genomes within incoming virus particles prior to virus replication. These findings not only highlight the sensitivity of cellular responses to low-level virus particle stimulation, but provide important insights into how nonreplicating virus vectors or synthetic lipid-based carriers used as clinical delivery vehicles activate innate immune responses. C ells defend themselves from viral infection by producing antiviral proteins, which cumulatively make them nonpermissive to virus replication (1). Large sets of antiviral proteins are induced by type I interferons (IFNs), and this response is critical for defense against viral infection (2, 3). IFN-␤ has no direct antiviral activity but signals induction of a set of IFN-stimulated genes (ISGs) encoding proteins with antiviral activity (4, 5). IFN-␤ is produced by a wide array of cells, and as it is the first IFN subtype produced in response to virus infection, many subsequen...

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Section: Discussioncontrasting

confidence: 50%

Membrane Perturbation-Associated Ca ²⁺ Signaling and Incoming Genome Sensing Are Required for the Host Response to Low-Level Enveloped Virus Particle Entry

Hare

Collins

Mukherjee

et al. 2016

J Virol

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“…The publication of crystal structures demonstrating the structural basis for CDN sensing through STING (64,65) and the identification of endogenous CDNs as signaling products produced by cyclic GMP-AMP synthase sensing of double-stranded DNA (9,10,66) have provided both a rationale and mechanistic guidance for the development of CDNs as adjuvants acting through type I IFNs and NF-κB activation in host cells. Indeed, several laboratories have recently demonstrated innate immune stimulatory (14,22,61) and adjuvant activities (63) with CDNs, particularly when applied Type I IFNs are signature downstream products of STING activation by CDNs in host cells (51,52), and NP-cdGMP led to direct induction of type I IFN and downstream target gene expression in the dLNs.…”

Section: Discussionmentioning

confidence: 99%

Nanoparticulate STING agonists are potent lymph node–targeted vaccine adjuvants

Hanson¹,

Crespo²,

Abraham³

et al. 2015

J. Clin. Invest.

324

310

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“…However, the DNA sensor involved in recognizing infection by adenovirus leading to early IRF3 activation has not been convincingly established. The recent identification of cyclic dinucleotide activation of STING (14)(15)(16)(17)(18) and the elegant discovery of cyclic-GMP-AMP synthase (cGAS) as a DNA sensor (19,20) provide an important bridge between DNA detection and downstream signaling. cGAS in complex with duplex DNA (21)(22)(23) leads to enzyme activation and production of a novel cyclic guanine-adenine dinucleotide (cGAMP) (24)(25)(26)(27) and STING activation.…”

mentioning

confidence: 99%

Adenovirus Detection by the cGAS/STING/TBK1 DNA Sensing Cascade

Lam

Stein

Falck-Pedersen

2014

J Virol

198

149

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T he human serotype 5 adenovirus (Ad5) is a nonenveloped linear double-stranded DNA virus associated with upper respiratory tract disease in humans. It has been extensively studied as a model for virus and host cell interactions. Replication-defective recombinant Ad5 vectors (rAdV) deleted in E1 and E3 coding domains have been characterized in gene therapy, vaccine, and oncolytic vector strategies in the murine model. Although nonpermissive for Ad5 replication, the murine model of rAdV infection provides a valuable resource for characterizing how the innate and adaptive immune response orchestrates an antiviral response to nonenveloped DNA viruses.Virus uptake by immune sentinel cells such as macrophage and dendritic cells is vital to initiating the antiviral immune response. In addition to antigen-presenting cells (APCs), other cell types, including endothelial cells or tissue-specific cells such as hepatocytes, when exposed to virus, also contribute to the host antiviral response. In vitro studies of isolated bone marrow-derived APCs or representative cell lines have revealed a cell-specific antiviral innate response, where activation of the type I interferon (IFN) cascade is a dominant feature (1-4). A valuable marker for early events in the antiviral recognition response is activation of the transcription factor interferon response factor 3 (IRF3). Following infection, cytosolic IRF3 undergoes phosphorylation as a primary response to adenovirus uptake. Activation occurs in a MyD88/TRIF-independent manner; it requires integrin-dependent endosomal entry, escape, and presentation of viral DNA to the cytosolic compartment (3).In rAdV-responsive murine cell lines, the STING/TBK1 cascade is required for IRF3 phosphorylation (5, 6). STING (7,8) functions as an adaptor linking DNA recognition signaling to activation of the TBK1 kinase. TBK1 activation (9) leads to C-terminal IRF3 phosphorylation, dimerization, and translocation to the nucleus (10, 11). In the nucleus, IRF3, in collaboration with additional transcription factors (NF-B and AP1), results in transcriptional activation of IRF3-responsive genes (including IFN-␤) (12). This sequence of events contributes to the primary antiviral response to adenovirus infection. The translation of primary response transcripts such as IFN-␤ leads to autocrine/paracrine secondary signaling. The combination of primary and secondary response functions leads to expression of a complete antiviral response, which is distinct for different cell types.Using various screening protocols, cell lines, and output assays, an extensive list of cytosolic DNA sensors, including DAI, RNA polymerase (Pol) III, IFI16, DDX41, and Aim 2, has been established (reviewed in reference 13). However, the DNA sensor involved in recognizing infection by adenovirus leading to early IRF3 activation has not been convincingly established. The recent identification of cyclic dinucleotide activation of STING (14-18) and the elegant discovery of cyclic-GMP-AMP synthase (cGAS) as a DNA sensor (19,20) provide an...

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Structure of STING bound to cyclic di-GMP reveals the mechanism of cyclic dinucleotide recognition by the immune system

Cited by 253 publications

References 23 publications

Membrane Perturbation-Associated Ca ²⁺ Signaling and Incoming Genome Sensing Are Required for the Host Response to Low-Level Enveloped Virus Particle Entry

Membrane Perturbation-Associated Ca ²⁺ Signaling and Incoming Genome Sensing Are Required for the Host Response to Low-Level Enveloped Virus Particle Entry

Nanoparticulate STING agonists are potent lymph node–targeted vaccine adjuvants

Adenovirus Detection by the cGAS/STING/TBK1 DNA Sensing Cascade

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Structure of STING bound to cyclic di-GMP reveals the mechanism of cyclic dinucleotide recognition by the immune system

Cited by 253 publications

References 23 publications

Membrane Perturbation-Associated Ca 2+ Signaling and Incoming Genome Sensing Are Required for the Host Response to Low-Level Enveloped Virus Particle Entry

Membrane Perturbation-Associated Ca 2+ Signaling and Incoming Genome Sensing Are Required for the Host Response to Low-Level Enveloped Virus Particle Entry

Nanoparticulate STING agonists are potent lymph node–targeted vaccine adjuvants

Adenovirus Detection by the cGAS/STING/TBK1 DNA Sensing Cascade

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Product

Resources

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Membrane Perturbation-Associated Ca ²⁺ Signaling and Incoming Genome Sensing Are Required for the Host Response to Low-Level Enveloped Virus Particle Entry

Membrane Perturbation-Associated Ca ²⁺ Signaling and Incoming Genome Sensing Are Required for the Host Response to Low-Level Enveloped Virus Particle Entry