STING specifies IRF3 phosphorylation by TBK1 in the cytosolic DNA signaling pathway

Yasuo Tanaka, Zhijian J. Chen

Research output: Contribution to journalArticle

334 Citations (Scopus)

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.

Original languageEnglish (US)
Article numberra20
JournalScience Signaling
Volume5
Issue number214
DOIs
StatePublished - Mar 6 2012

Fingerprint

Interferon Regulatory Factor-3
Phosphorylation
Phosphotransferases
Genes
Interferon Type I
Proteins
Scaffolds
Endoplasmic Reticulum
Interferons
Transcription Factors
Chemical activation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

STING specifies IRF3 phosphorylation by TBK1 in the cytosolic DNA signaling pathway. / Tanaka, Yasuo; Chen, Zhijian J.

In: Science Signaling, Vol. 5, No. 214, ra20, 06.03.2012.

Research output: Contribution to journalArticle

@article{fc61b60fef01420eaebf30caf87182ad,
title = "STING specifies IRF3 phosphorylation by TBK1 in the cytosolic DNA signaling pathway",
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.",
author = "Yasuo Tanaka and Chen, {Zhijian J.}",
year = "2012",
month = "3",
day = "6",
doi = "10.1126/scisignal.2002521",
language = "English (US)",
volume = "5",
journal = "Science Signaling",
issn = "1937-9145",
publisher = "American Association for the Advancement of Science",
number = "214",

}

TY - JOUR

T1 - STING specifies IRF3 phosphorylation by TBK1 in the cytosolic DNA signaling pathway

AU - Tanaka, Yasuo

AU - Chen, Zhijian J.

PY - 2012/3/6

Y1 - 2012/3/6

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84857937262&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84857937262&partnerID=8YFLogxK

U2 - 10.1126/scisignal.2002521

DO - 10.1126/scisignal.2002521

M3 - Article

VL - 5

JO - Science Signaling

JF - Science Signaling

SN - 1937-9145

IS - 214

M1 - ra20

ER -