STING Specifies IRF3 Phosphorylation by TBK1 in the Cytosolic DNA Signaling Pathway

Sci. Signal., 6 March 2012
Vol. 5, Issue 214, p. ra20
DOI: 10.1126/scisignal.2002521

STING Specifies IRF3 Phosphorylation by TBK1 in the Cytosolic DNA Signaling Pathway

  1. Yasuo Tanaka1 and
  2. Zhijian J. Chen1,2,*
  1. 1Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390–9148, USA.
  2. 2Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, TX 75390–9148, USA.
  1. *To whom correspondence should be addressed. E-mail: zhijian.chen{at}utsouthwestern.edu

Abstract

Cytosolic double-stranded DNA (dsDNA) stimulates the production of type I interferon (IFN) through the endoplasmic reticulum (ER)–resident adaptor protein STING (stimulator of IFN genes), which activates the transcription factor interferon regulatory factor 3 (IRF3); however, how STING activates IRF3 is unclear. Here, we showed that STING stimulates phosphorylation of IRF3 by the kinase TBK1 (TANK-binding kinase 1) in an in vitro reconstitution system. With this system, we identified a carboxyl-terminal region of STING that was both necessary and sufficient to activate TBK1 and stimulate the phosphorylation of IRF3. We also found that STING interacted with both TBK1 and IRF3 and that mutations in STING that selectively disrupted its binding to IRF3 abrogated phosphorylation of IRF3 without impairing the activation of TBK1. These results suggest that STING functions as a scaffold protein to specify and promote the phosphorylation of IRF3 by TBK1. This scaffolding function of STING (and possibly of other adaptor proteins) may explain why IRF3 is activated in only a subset of signaling pathways that activate TBK1.

Citation:

Y. Tanaka and Z. J. Chen, STING Specifies IRF3 Phosphorylation by TBK1 in the Cytosolic DNA Signaling Pathway. Sci. Signal. 5, ra20 (2012).

Dengue Virus NS Proteins Inhibit RIG-I/MAVS Signaling by Blocking TBK1/IRF3 Phosphorylation: Dengue Virus Serotype 1 NS4A Is a Unique Interferon-Regulating Virulence Determinant
N. A. Dalrymple, V. Cimica, and E. R. Mackow
mBio 6, e00553-15-e00553-15 (1 July 2015)

UV Light Potentiates STING (Stimulator of Interferon Genes)-dependent Innate Immune Signaling through Deregulation of ULK1 (Unc51-like Kinase 1)
M. G. Kemp, L. A. Lindsey-Boltz, and A. Sancar
J Biol Chem 290, 12184-12194 (8 May 2015)

Activation and Regulation of DNA-Driven Immune Responses
S. R. Paludan, L. A. Lindsey-Boltz, and A. Sancar
Microbiol. Mol. Biol. Rev. 79, 225-241 (29 April 2015)

STING agonist formulated cancer vaccines can cure established tumors resistant to PD-1 blockade
J. Fu, D. B. Kanne, M. Leong, L. H. Glickman, S. M. McWhirter, E. Lemmens, K. Mechette, J. J. Leong, P. Lauer, W. Liu et al.
Sci Transl Med 7, 283ra52-283ra52 (15 April 2015)

cGAS and Ifi204 Cooperate To Produce Type I IFNs in Response to Francisella Infection
K. M. Storek, N. A. Gertsvolf, M. B. Ohlson, and D. M. Monack
J. Immunol. 194, 3236-3245 (1 April 2015)

International Union of Basic and Clinical Pharmacology. XCVI. Pattern Recognition Receptors in Health and Disease
C. E. Bryant, S. Orr, B. Ferguson, M. F. Symmons, J. P. Boyle, and T. P. Monie
Pharmacol. Rev. 67, 462-504 (31 March 2015)

Phosphorylation of innate immune adaptor proteins MAVS, STING, and TRIF induces IRF3 activation
S. Liu, X. Cai, J. Wu, Q. Cong, X. Chen, T. Li, F. Du, J. Ren, Y.-T. Wu, N. V. Grishin et al.
Science 347, aaa2630-aaa2630 (13 March 2015)

Emerging Principles Governing Signal Transduction by Pattern-Recognition Receptors
J. C. Kagan, and G. M. Barton
Cold Spring Harb. Perspect. Biol. 7, a016253-a016253 (1 March 2015)

The functional interactome of PYHIN immune regulators reveals IFIX is a sensor of viral DNA
B. A. Diner, T. Li, T. M. Greco, M. S. Crow, J. A. Fuesler, J. Wang, and I. M. Cristea
Mol Syst Biol 11, 787-787 (27 February 2015)

Hepatitis B Virus Polymerase Disrupts K63-Linked Ubiquitination of STING To Block Innate Cytosolic DNA-Sensing Pathways
Y. Liu, J. Li, J. Chen, Y. Li, W. Wang, X. Du, W. Song, W. Zhang, L. Lin, Z. Yuan et al.
J. Virol. 89, 2287-2300 (15 February 2015)

Positive Feedback Regulation of Type I IFN Production by the IFN-Inducible DNA Sensor cGAS
F. Ma, B. Li, S.-y. Liu, S. S. Iyer, Y. Yu, A. Wu, and G. Cheng
J. Immunol. 194, 1545-1554 (15 February 2015)

Microbial Sensing by Toll-Like Receptors and Intracellular Nucleic Acid Sensors
S. Pandey, T. Kawai, and S. Akira
Cold Spring Harb. Perspect. Biol. 7, a016246-a016246 (1 January 2015)

IKK{beta} is an IRF5 kinase that instigates inflammation
J. Ren, X. Chen, and Z. J. Chen
Proc. Natl. Acad. Sci. USA 111, 17438-17443 (9 December 2014)

Stimulation of Innate Immunity by In Vivo Cyclic di-GMP Synthesis Using Adenovirus
B. J. Koestler, S. S. Seregin, D. P. W. Rastall, Y. A. Aldhamen, S. Godbehere, A. Amalfitano, and C. M. Waters
CVI 21, 1550-1559 (1 November 2014)

Identification of a negative feedback loop between cyclic di-GMP-induced levels of IFI16 and p202 cytosolic DNA sensors and STING
R. Panchanathan, H. Liu, D. Xin, and D. Choubey
Innate Immunity 20, 751-759 (1 October 2014)

Listeria monocytogenes induces IFN{beta} expression through an IFI16-, cGAS- and STING-dependent pathway
K. Hansen, T. Prabakaran, A. Laustsen, S. E. Jorgensen, S. H. Rahbaek, S. B. Jensen, R. Nielsen, J. H. Leber, T. Decker, K. A. Horan et al.
EMBO J. 33, 1654-1666 (1 August 2014)

DNA Degradation and Its Defects
K. Kawane, K. Motani, and S. Nagata
Cold Spring Harb. Perspect. Biol. 6, a016394-a016394 (1 June 2014)

Cytosolic-DNA-Mediated, STING-Dependent Proinflammatory Gene Induction Necessitates Canonical NF-{kappa}B Activation through TBK1
T. Abe, and G. N. Barber
J. Virol. 88, 5328-5341 (15 May 2014)

Age-Enhanced Endoplasmic Reticulum Stress Contributes to Increased Atg9A Inhibition of STING-Mediated IFN-{beta} Production during Streptococcus pneumoniae Infection
D. N. Mitzel, V. Lowry, A. C. Shirali, Y. Liu, and H. W. Stout-Delgado
J. Immunol. 192, 4273-4283 (1 May 2014)

An Alternative Splicing Isoform of MITA Antagonizes MITA-Mediated Induction of Type I IFNs
H. Chen, R. Pei, W. Zhu, R. Zeng, Y. Wang, Y. Wang, M. Lu, and X. Chen
J. Immunol. 192, 1162-1170 (1 February 2014)

siRNA enhances DNA-mediated interferon lambda-1 response through crosstalk between RIG-I and IFI16 signalling pathway
H. Sui, M. Zhou, Q. Chen, H. C. Lane, and T. Imamichi
Nucleic Acids Res 42, 583-598 (1 January 2014)

Dengue Virus Subverts the Interferon Induction Pathway via NS2B/3 Protease-I{kappa}B Kinase {varepsilon} Interaction
Y. I. Anglero-Rodriguez, P. Pantoja, C. A. Sariol, and W. R. Waters
CVI 21, 29-38 (1 January 2014)

The Viral Ubiquitin Ligase ICP0 Is neither Sufficient nor Necessary for Degradation of the Cellular DNA Sensor IFI16 during Herpes Simplex Virus 1 Infection
D. Cuchet-Lourenco, G. Anderson, E. Sloan, A. Orr, and R. D. Everett
J. Virol. 87, 13422-13432 (15 December 2013)

STING-IRF3 pathway links endoplasmic reticulum stress with hepatocyte apoptosis in early alcoholic liver disease
J. Petrasek, A. Iracheta-Vellve, T. Csak, A. Satishchandran, K. Kodys, E. A. Kurt-Jones, K. A. Fitzgerald, and G. Szabo
Proc. Natl. Acad. Sci. USA 110, 16544-16549 (8 October 2013)

IFIT5 potentiates anti-viral response through enhancing innate immune signaling pathways
B. Zhang, X. Liu, W. Chen, and L. Chen
Acta Biochim Biophys Sin 45, 867-874 (1 October 2013)

Enhanced optineurin E50K-TBK1 interaction evokes protein insolubility and initiates familial primary open-angle glaucoma
Y. Minegishi, D. Iejima, H. Kobayashi, Z.-L. Chi, K. Kawase, T. Yamamoto, T. Seki, S. Yuasa, K. Fukuda, T. Iwata et al.
Hum Mol Genet 22, 3559-3567 (1 September 2013)

Human DEAD Box Helicase 3 Couples I{kappa}B Kinase {varepsilon} to Interferon Regulatory Factor 3 Activation
L. Gu, A. Fullam, R. Brennan, and M. Schroder
Mol. Cell. Biol. 33, 2004-2015 (15 May 2013)

Mouse, but not Human STING, Binds and Signals in Response to the Vascular Disrupting Agent 5,6-Dimethylxanthenone-4-Acetic Acid
J. Conlon, D. L. Burdette, S. Sharma, N. Bhat, M. Thompson, Z. Jiang, V. A. K. Rathinam, B. Monks, T. Jin, T. S. Xiao et al.
J. Immunol. 190, 5216-5225 (15 May 2013)

STING-Dependent Recognition of Cyclic di-AMP Mediates Type I Interferon Responses during Chlamydia trachomatis Infection
J. R. Barker, B. J. Koestler, V. K. Carpenter, D. L. Burdette, C. M. Waters, R. E. Vance, and R. H. Valdivia
mBio 4, e00018-13-e00018-13 (30 April 2013)

Proteasomal Degradation of Herpes Simplex Virus Capsids in Macrophages Releases DNA to the Cytosol for Recognition by DNA Sensors
K. A. Horan, K. Hansen, M. R. Jakobsen, C. K. Holm, S. Soby, L. Unterholzner, M. Thompson, J. A. West, M. B. Iversen, S. B. Rasmussen et al.
J. Immunol. 190, 2311-2319 (1 March 2013)

Cyclic GMP-AMP Synthase Is a Cytosolic DNA Sensor That Activates the Type I Interferon Pathway
L. Sun, J. Wu, F. Du, X. Chen, and Z. J. Chen
Science 339, 786-791 (15 February 2013)

Cyclic GMP-AMP Is an Endogenous Second Messenger in Innate Immune Signaling by Cytosolic DNA
J. Wu, L. Sun, X. Chen, F. Du, H. Shi, C. Chen, and Z. J. Chen
Science 339, 826-830 (15 February 2013)

The IRF Family Transcription Factors at the Interface of Innate and Adaptive Immune Responses
H. Ikushima, H. Negishi, and T. Taniguchi
Cold Spring Harb Symp Quant Biol 78, 105-116 (1 January 2013)

MAVS recruits multiple ubiquitin E3 ligases to activate antiviral signaling cascades
S. Liu, J. Chen, X. Cai, J. Wu, X. Chen, Y.-T. Wu, L. Sun, and Z. J. Chen
elife 2, e00785-e00785 (1 January 2013)

ORF45 of Kaposi's Sarcoma-Associated Herpesvirus Inhibits Phosphorylation of Interferon Regulatory Factor 7 by IKK{varepsilon} and TBK1 as an Alternative Substrate
Q. Liang, B. Fu, F. Wu, X. Li, Y. Yuan, and F. Zhu
J. Virol. 86, 10162-10172 (15 September 2012)

The STING in the Tail for Cytosolic DNA-Dependent Activation of IRF3
A. Bowie, B. Fu, F. Wu, X. Li, Y. Yuan, and F. Zhu
Sci Signal 5, pe9-pe9 (6 March 2012)

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882