Structural basis of STING binding with and phosphorylation by TBK1
Open accessAbstract
The invasion of mammalian cytoplasm by microbial DNA from infectious pathogens or by self DNA from the nucleus or mitochondria represents a danger signal that alerts the host immune system 1. Cyclic GMP-AMP synthase (cGAS) is a sensor of cytoplasmic DNA that activates the type-I interferon pathway 2. On binding to DNA, cGAS is activated to catalyse the synthesis of cyclic GMP-AMP (cGAMP) from GTP and ATP 3. cGAMP functions as a second messenger that binds to and activates stimulator of interferon genes (STING)3-9. STING then recruits and activates tank-binding kinase 1 (TBK1), which phosphorylates STING and the transcription factor IRF3 to induce type-I interferons and other cytokines 10,11. However, how cGAMP-bound STING activates TBK1 and IRF3 is not understood. Here we present the cryo-electron microscopy structure of human TBK1 in complex with cGAMP-bound, full-length chicken STING. The structure reveals that the C-terminal tail of STING adopts a β-strand-like conformation and inserts into a groove between the kinase domain of one TBK1 subunit and the scaffold and dimerization domain of the second subunit in the TBK1 dimer. In this binding mode, the phosphorylation site Ser366 in the STING tail cannot reach the kinase-domain active site of bound TBK1, which suggests that STING phosphorylation by TBK1 requires the oligomerization of both proteins. Mutational analyses validate the interaction mode between TBK1 and STING and support a model in which high-order oligomerization of STING and TBK1, induced by cGAMP, leads to STING phosphorylation by TBK1.
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On reaching the Golgi, STING recruits TBK1, which activates IRF3 and NF-κB, leading to expression of ISGs, type I interferons and other inflammatory cytokines.
“On reaching the Golgi compartment, STING then recruits TBK1, which then activates IRF3 and NF-κB, leading to the expression of interferon-stimulated genes (ISGs), type I interferons and other inflammatory cytokines (e.g., IL-6, IL-12).393–396”
From Role of neuroinflammation in neurodegeneration development (2023) · cites this paper as [393]
The Abstract reports that STING recruits TBK1, which phosphorylates IRF3 to induce type-I interferons and other cytokines; Extended Data Fig. 7 also describes IRF3 initiating IFN-gene transcription. The paper places STING–TBK1 puncta in a perinuclear compartment, not specifically the Golgi, and the passages do not establish NF-κB activation, ISG expression, or IL-6 and IL-12 as outcomes. (LLM, conf=0.87)
Experimental — accuracy not yet measuredLevels of phosphorylated TBK1 (Ser172) are increased in aged mouse brains.
“The levels of phosphorylated TBK1 (Ser172; pTBK1)—the major kinase responsible for signal transduction downstream of STING19,20—were increased in aged mouse brains (Fig. 2d).”
From cGAS–STING drives ageing-related inflammation and neurodegeneration (2023) · cites this paper as [19]
cGAMP-induced STING oligomerization recruits and activates TBK1.
“cGAMP-induced STING oligomerization provides a signaling platform for recruiting and activating TANK-binding kinase (TBK1) (Zhang et al., 2019).”
From Cytoplasmic DNA: sources, sensing, and role in aging and disease (2021) · cites this paper as Zhang et al., 2019
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- Authors
- Zhang, C., Shang, G., Gui, X., Zhang, X., Bai, X.-C., & Chen, Z. J.
- Affiliations
- The University of Texas Southwestern Medical Center (US)
- Howard Hughes Medical Institute (US)
- Corresponding author
- Zhang, Xuewu
- Bai, Xiao-chen
- Chen, Zhijian J.
- Venue
- Nature
- Published
- 2019-03-06
- Volume
- 567
- Issue
- 7748
- Pages
- 394-398
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC6862768
- OpenAlex
- W2919704769
- Cited by
- 1,095
- References
- 27
- Citations by year
- 2026: 177 · 2025: 212 · 2024: 198 · 2023: 168 · 2022: 135 · 2021: 102 · 2020: 79 · 2019: 24
- FWCI
- 34.30
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- Funders
- Howard Hughes Medical Institute
- Welch Foundation
- University of Texas Southwestern Medical Center
- National Institutes of Health
- National Institute of General Medical Sciences (R01GM088197, R35GM130289)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › interferon and immune responses
- Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Keywords
- STING, TBK1, cGAMP, STING phosphorylation, protein oligomerization, cytosolic DNA sensing, type I interferon, IRF3, cGAS, interferon-stimulated genes, protein complex structure
- MeSH terms
- Animals, Binding Sites, Chickens, HeLa Cells, Humans, Membrane Proteins, Models, Molecular, Mutation, Nucleotides, Cyclic, Phosphorylation, Protein Binding, Protein Serine-Threonine Kinases, Catalytic Domain, Cryoelectron Microscopy, Protein Multimerization, HEK293 Cells
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