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Title: A conserved PLPLRT/SD motif of STING mediates the recruitment and activation of TBK1

Abstract

Nucleic acids from bacteria or viruses induce potent immune responses in infected cells. The detection of pathogen-derived nucleic acids is a central strategy by which the host senses infection and initiates protective immune responses. Cyclic GMP-AMP synthase (cGAS) is a double-stranded DNA sensor. It catalyses the synthesis of cyclic GMP-AMP (cGAMP), which stimulates the induction of type I interferons through the STING-TBK1-IRF-3 signalling axis. STING oligomerizes after binding of cGAMP, leading to the recruitment and activation of the TBK1 kinase. The IRF-3 transcription factor is then recruited to the signalling complex and activated by TBK1. Phosphorylated IRF-3 translocates to the nucleus and initiates the expression of type I interferons. However, the precise mechanisms that govern activation of STING by cGAMP and subsequent activation of TBK1 by STING remain unclear. Here we show that a conserved PLPLRT/SD motif within the C-terminal tail of STING mediates the recruitment and activation of TBK1. Crystal structures of TBK1 bound to STING reveal that the PLPLRT/SD motif binds to the dimer interface of TBK1. Cell-based studies confirm that the direct interaction between TBK1 and STING is essential for induction of IFNβ after cGAMP stimulation. Moreover, we show that full-length STING oligomerizes after it binds cGAMP,more » and highlight this as an essential step in the activation of STING-mediated signalling. Furthermore, these findings provide a structural basis for the development of STING agonists and antagonists for the treatment of cancer and autoimmune disorders.« less

Authors:
 [1];  [1];  [1];  [1];  [2];  [1];  [3];  [3];  [3];  [4];  [4];  [1];  [1];  [1];  [3];  [3];  [3];  [1]
  1. Texas A & M Univ., College Station, TX (United States)
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
  3. Texas A & M Univ. Health Science Center, College Station, TX (United States)
  4. Florida State Univ., Tallahassee, FL (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1577334
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Nature (London)
Additional Journal Information:
Journal Name: Nature (London); Journal Volume: 569; Journal Issue: 7758; Journal ID: ISSN 0028-0836
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Zhao, Baoyu, Du, Fenglei, Xu, Pengbiao, Shu, Chang, Sankaran, Banumathi, Bell, Samantha L., Liu, Mengmeng, Lei, Yuanjiu, Gao, Xinsheng, Fu, Xiaofeng, Zhu, Fanxiu, Liu, Yang, Laganowsky, Arthur, Zheng, Xueyun, Ji, Jun -Yuan, West, A. Phillip, Watson, Robert O., and Li, Pingwei. A conserved PLPLRT/SD motif of STING mediates the recruitment and activation of TBK1. United States: N. p., 2019. Web. doi:10.1038/s41586-019-1228-x.
Zhao, Baoyu, Du, Fenglei, Xu, Pengbiao, Shu, Chang, Sankaran, Banumathi, Bell, Samantha L., Liu, Mengmeng, Lei, Yuanjiu, Gao, Xinsheng, Fu, Xiaofeng, Zhu, Fanxiu, Liu, Yang, Laganowsky, Arthur, Zheng, Xueyun, Ji, Jun -Yuan, West, A. Phillip, Watson, Robert O., & Li, Pingwei. A conserved PLPLRT/SD motif of STING mediates the recruitment and activation of TBK1. United States. doi:10.1038/s41586-019-1228-x.
Zhao, Baoyu, Du, Fenglei, Xu, Pengbiao, Shu, Chang, Sankaran, Banumathi, Bell, Samantha L., Liu, Mengmeng, Lei, Yuanjiu, Gao, Xinsheng, Fu, Xiaofeng, Zhu, Fanxiu, Liu, Yang, Laganowsky, Arthur, Zheng, Xueyun, Ji, Jun -Yuan, West, A. Phillip, Watson, Robert O., and Li, Pingwei. Wed . "A conserved PLPLRT/SD motif of STING mediates the recruitment and activation of TBK1". United States. doi:10.1038/s41586-019-1228-x.
@article{osti_1577334,
title = {A conserved PLPLRT/SD motif of STING mediates the recruitment and activation of TBK1},
author = {Zhao, Baoyu and Du, Fenglei and Xu, Pengbiao and Shu, Chang and Sankaran, Banumathi and Bell, Samantha L. and Liu, Mengmeng and Lei, Yuanjiu and Gao, Xinsheng and Fu, Xiaofeng and Zhu, Fanxiu and Liu, Yang and Laganowsky, Arthur and Zheng, Xueyun and Ji, Jun -Yuan and West, A. Phillip and Watson, Robert O. and Li, Pingwei},
abstractNote = {Nucleic acids from bacteria or viruses induce potent immune responses in infected cells. The detection of pathogen-derived nucleic acids is a central strategy by which the host senses infection and initiates protective immune responses. Cyclic GMP-AMP synthase (cGAS) is a double-stranded DNA sensor. It catalyses the synthesis of cyclic GMP-AMP (cGAMP), which stimulates the induction of type I interferons through the STING-TBK1-IRF-3 signalling axis. STING oligomerizes after binding of cGAMP, leading to the recruitment and activation of the TBK1 kinase. The IRF-3 transcription factor is then recruited to the signalling complex and activated by TBK1. Phosphorylated IRF-3 translocates to the nucleus and initiates the expression of type I interferons. However, the precise mechanisms that govern activation of STING by cGAMP and subsequent activation of TBK1 by STING remain unclear. Here we show that a conserved PLPLRT/SD motif within the C-terminal tail of STING mediates the recruitment and activation of TBK1. Crystal structures of TBK1 bound to STING reveal that the PLPLRT/SD motif binds to the dimer interface of TBK1. Cell-based studies confirm that the direct interaction between TBK1 and STING is essential for induction of IFNβ after cGAMP stimulation. Moreover, we show that full-length STING oligomerizes after it binds cGAMP, and highlight this as an essential step in the activation of STING-mediated signalling. Furthermore, these findings provide a structural basis for the development of STING agonists and antagonists for the treatment of cancer and autoimmune disorders.},
doi = {10.1038/s41586-019-1228-x},
journal = {Nature (London)},
number = 7758,
volume = 569,
place = {United States},
year = {2019},
month = {5}
}

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Works referenced in this record:

STING Specifies IRF3 Phosphorylation by TBK1 in the Cytosolic DNA Signaling Pathway
journal, March 2012


Cyclic GMP-AMP Containing Mixed Phosphodiester Linkages Is An Endogenous High-Affinity Ligand for STING
journal, July 2013


Triggering the Interferon Antiviral Response Through an IKK-Related Pathway
journal, April 2003


Regulation and function of the cGAS–STING pathway of cytosolic DNA sensing
journal, September 2016

  • Chen, Qi; Sun, Lijun; Chen, Zhijian J.
  • Nature Immunology, Vol. 17, Issue 10
  • DOI: 10.1038/ni.3558

Virus-Dependent Phosphorylation of the IRF-3 Transcription Factor Regulates Nuclear Translocation, Transactivation Potential, and Proteasome-Mediated Degradation
journal, May 1998

  • Lin, Rongtuan; Heylbroeck, Christophe; Pitha, Paula M.
  • Molecular and Cellular Biology, Vol. 18, Issue 5
  • DOI: 10.1128/MCB.18.5.2986

Structural Insights into the Functions of TBK1 in Innate Antimicrobial Immunity
journal, July 2013


Structural basis for concerted recruitment and activation of IRF-3 by innate immune adaptor proteins
journal, June 2016

  • Zhao, Baoyu; Shu, Chang; Gao, Xinsheng
  • Proceedings of the National Academy of Sciences, Vol. 113, Issue 24
  • DOI: 10.1073/pnas.1603269113

RIG-I-like receptors: cytoplasmic sensors for non-self RNA: RIG-I-like receptors
journal, August 2011


Coot model-building tools for molecular graphics
journal, November 2004

  • Emsley, Paul; Cowtan, Kevin
  • Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12, p. 2126-2132
  • DOI: 10.1107/S0907444904019158

Cyclic GMP-AMP Synthase Is a Cytosolic DNA Sensor That Activates the Type I Interferon Pathway
journal, December 2012


Cryo-EM structures of STING reveal its mechanism of activation by cyclic GMP–AMP
journal, March 2019


The Innate Immune DNA Sensor cGAS Produces a Noncanonical Cyclic Dinucleotide that Activates Human STING
journal, May 2013


PHENIX: a comprehensive Python-based system for macromolecular structure solution
journal, January 2010

  • Adams, Paul D.; Afonine, Pavel V.; Bunkóczi, Gábor
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2, p. 213-221
  • DOI: 10.1107/S0907444909052925

Innate Immune Sensing and Signaling of Cytosolic Nucleic Acids
journal, March 2014


Genome-Scale CRISPR-Cas9 Knockout Screening in Human Cells
journal, December 2013

  • Shalem, Ophir; Sanjana, Neville E.; Hartenian, Ella
  • Science, Vol. 343, Issue 6166, p. 84-87
  • DOI: 10.1126/science.1247005

STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling
journal, August 2008


STING is a direct innate immune sensor of cyclic di-GMP
journal, September 2011

  • Burdette, Dara L.; Monroe, Kathryn M.; Sotelo-Troha, Katia
  • Nature, Vol. 478, Issue 7370
  • DOI: 10.1038/nature10429

Innate immune DNA sensing pathways: STING, AIMII and the regulation of interferon production and inflammatory responses
journal, February 2011


Integrating macromolecular X-ray diffraction data with the graphical user interface iMosflm
journal, June 2017

  • Powell, Harold R.; Battye, T. Geoff G.; Kontogiannis, Luke
  • Nature Protocols, Vol. 12, Issue 7
  • DOI: 10.1038/nprot.2017.037

cGAS produces a 2′-5′-linked cyclic dinucleotide second messenger that activates STING
journal, May 2013

  • Ablasser, Andrea; Goldeck, Marion; Cavlar, Taner
  • Nature, Vol. 498, Issue 7454
  • DOI: 10.1038/nature12306

Immune Sensing of DNA
journal, May 2013


Recognition of Endogenous Nucleic Acids by the Innate Immune System
journal, April 2016


Experimental phasing with SHELXC / D / E : combining chain tracing with density modification
journal, March 2010

  • Sheldrick, George M.
  • Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 4
  • DOI: 10.1107/S0907444909038360

Structure of STING bound to cyclic di-GMP reveals the mechanism of cyclic dinucleotide recognition by the immune system
journal, June 2012

  • Shu, Chang; Yi, Guanghui; Watts, Tylan
  • Nature Structural & Molecular Biology, Vol. 19, Issue 7
  • DOI: 10.1038/nsmb.2331

STING and the innate immune response to nucleic acids in the cytosol
journal, December 2012

  • Burdette, Dara L.; Vance, Russell E.
  • Nature Immunology, Vol. 14, Issue 1
  • DOI: 10.1038/ni.2491

IKKε and TBK1 are essential components of the IRF3 signaling pathway
journal, April 2003

  • Fitzgerald, Katherine A.; McWhirter, Sarah M.; Faia, Kerrie L.
  • Nature Immunology, Vol. 4, Issue 5
  • DOI: 10.1038/ni921

Phosphorylation of innate immune adaptor proteins MAVS, STING, and TRIF induces IRF3 activation
journal, January 2015


STING regulates intracellular DNA-mediated, type I interferon-dependent innate immunity
journal, September 2009

  • Ishikawa, Hiroki; Ma, Zhe; Barber, Glen N.
  • Nature, Vol. 461, Issue 7265
  • DOI: 10.1038/nature08476

Cyclic GMP-AMP Is an Endogenous Second Messenger in Innate Immune Signaling by Cytosolic DNA
journal, December 2012