Home LiteratureArticle Details
PMID: 23722159 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Structural mechanism of cytosolic DNA sensing by cGAS.

Nature ·Vol. 498 ·No. 7454 ·2013-06-20 ·Pages 332-7

Civril F, Deimling T, de Oliveira Mann CC, Ablasser A, Moldt M, Witte G, Hornung V, Hopfner KP

Abstract

Cytosolic DNA arising from intracellular bacterial or viral infections is a powerful pathogen-associated molecular pattern (PAMP) that leads to innate immune host defence by the production of type I interferon and inflammatory cytokines. Recognition of cytosolic DNA by the recently discovered cyclic-GMP-AMP (cGAMP) synthase (cGAS) induces the production of cGAMP to activate the stimulator of interferon genes (STING). Here we report the crystal structure of cGAS alone and in complex with DNA, ATP and GTP along with functional studies. Our results explain the broad DNA sensing specificity of cGAS, show how cGAS catalyses dinucleotide formation and indicate activation by a DNA-induced structural switch. cGAS possesses a remarkable structural similarity to the antiviral cytosolic double-stranded RNA sensor 2'-5'oligoadenylate synthase (OAS1), but contains a unique zinc thumb that recognizes B-form double-stranded DNA. Our results mechanistically unify dsRNA and dsDNA innate immune sensing by OAS1 and cGAS nucleotidyl transferases.

MeSH Terms
Adenosine Triphosphate/chemistry,metabolism Animals Base Sequence Catalytic Domain Crystallography, X-Ray Cytosol DNA/chemistry,metabolism,pharmacology Guanosine Triphosphate/chemistry,metabolism HEK293 Cells Humans Membrane Proteins/genetics,metabolism Mice Models, Biological Models, Molecular Mutation Nucleotidyltransferases/chemistry,genetics,metabolism Protein Conformation/drug effects Structure-Activity Relationship Substrate Specificity Swine Uridine Triphosphate/chemistry,metabolism Zinc/chemistry,metabolism
Chemicals
Membrane Proteins Sting1 protein, mouse Guanosine Triphosphate Adenosine Triphosphate DNA Nucleotidyltransferases cGAS protein, human Zinc Uridine Triphosphate
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Civril Filiz
Department of Biochemistry and Gene Center, Ludwig-Maximilians-University, 81377 Munich, Germany.
Deimling Tobias
de Oliveira Mann Carina C
Ablasser Andrea
Moldt Manuela
Witte Gregor
Hornung Veit
Hopfner Karl-Peter
References (39)
39 references, click to expand
  1. AIM2 activates the inflammasome and cell death in response to cytoplasmic DNA.
    Nature. 2009 Mar 26;458(7237):509-13 PMID: 19158676
  2. Pattern recognition receptors and inflammation.
    Cell. 2010 Mar 19;140(6):805-20 PMID: 20303872
  3. Crystal structure of the 2'-specific and double-stranded RNA-activated interferon-induced antiviral protein 2'-5'-oligoadenylate synthetase.
    Mol Cell. 2003 Nov;12(5):1173-85 PMID: 14636576
  4. Aspartate-glutamate-alanine-histidine box motif (DEAH)/RNA helicase A helicases sense microbial DNA in human plasmacytoid dendritic cells.
    Proc Natl Acad Sci U S A. 2010 Aug 24;107(34):15181-6 PMID: 20696886
  5. Automated structure solution with autoSHARP.
    Methods Mol Biol. 2007;364:215-30 PMID: 17172768
  6. STING recognition of cytoplasmic DNA instigates cellular defense.
    Mol Cell. 2013 Apr 11;50(1):5-15 PMID: 23478444
  7. Structural biochemistry of a bacterial checkpoint protein reveals diadenylate cyclase activity regulated by DNA recombination intermediates.
    Mol Cell. 2008 Apr 25;30(2):167-78 PMID: 18439896
  8. Nucleic acid recognition receptors in autoimmunity.
    Handb Exp Pharmacol. 2008;(183):129-51 PMID: 18071658
  9. Comprehensive classification of nucleotidyltransferase fold proteins: identification of novel families and their representatives in human.
    Nucleic Acids Res. 2009 Dec;37(22):7701-14 PMID: 19833706
  10. Species-specific detection of the antiviral small-molecule compound CMA by STING.
    EMBO J. 2013 May 15;32(10):1440-50 PMID: 23604073
  11. IFI16 is an innate immune sensor for intracellular DNA.
    Nat Immunol. 2010 Nov;11(11):997-1004 PMID: 20890285
  12. DAI (DLM-1/ZBP1) is a cytosolic DNA sensor and an activator of innate immune response.
    Nature. 2007 Jul 26;448(7152):501-5 PMID: 17618271
  13. ARP/wARP's model-building algorithms. I. The main chain.
    Acta Crystallogr D Biol Crystallogr. 2002 Jun;58(Pt 6 Pt 2):968-75 PMID: 12037299
  14. Structural basis for cytosolic double-stranded RNA surveillance by human oligoadenylate synthetase 1.
    Proc Natl Acad Sci U S A. 2013 Jan 29;110(5):1652-7 PMID: 23319625
  15. The helicase DDX41 senses intracellular DNA mediated by the adaptor STING in dendritic cells.
    Nat Immunol. 2011 Sep 04;12(10):959-65 PMID: 21892174
  16. Cytosolic surveillance and antiviral immunity.
    Curr Opin Virol. 2011 Dec;1(6):455-62 PMID: 22440909
  17. c-di-AMP secreted by intracellular Listeria monocytogenes activates a host type I interferon response.
    Science. 2010 Jun 25;328(5986):1703-5 PMID: 20508090
  18. AIM2 recognizes cytosolic dsDNA and forms a caspase-1-activating inflammasome with ASC.
    Nature. 2009 Mar 26;458(7237):514-8 PMID: 19158675
  19. The Buccaneer software for automated model building. 1. Tracing protein chains.
    Acta Crystallogr D Biol Crystallogr. 2006 Sep;62(Pt 9):1002-11 PMID: 16929101
  20. Refinement of severely incomplete structures with maximum likelihood in BUSTER-TNT.
    Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2210-21 PMID: 15572774
  21. The C-terminal regulatory domain is the RNA 5'-triphosphate sensor of RIG-I.
    Mol Cell. 2008 Feb 1;29(2):169-79 PMID: 18243112
  22. STING regulates intracellular DNA-mediated, type I interferon-dependent innate immunity.
    Nature. 2009 Oct 8;461(7265):788-92 PMID: 19776740
  23. Coot: model-building tools for molecular graphics.
    Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32 PMID: 15572765
  24. Cyclic GMP-AMP synthase is a cytosolic DNA sensor that activates the type I interferon pathway.
    Science. 2013 Feb 15;339(6121):786-91 PMID: 23258413
  25. UTP-bound and Apo structures of a minimal RNA uridylyltransferase.
    J Mol Biol. 2007 Feb 23;366(3):882-99 PMID: 17189640
  26. A host type I interferon response is induced by cytosolic sensing of the bacterial second messenger cyclic-di-GMP.
    J Exp Med. 2009 Aug 31;206(9):1899-911 PMID: 19652017
  27. PHENIX: a comprehensive Python-based system for macromolecular structure solution.
    Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21 PMID: 20124702
  28. Cyclic GMP-AMP is an endogenous second messenger in innate immune signaling by cytosolic DNA.
    Science. 2013 Feb 15;339(6121):826-30 PMID: 23258412
  29. Cytosolic DNA sensors regulating type I interferon induction.
    Trends Immunol. 2011 Dec;32(12):574-81 PMID: 21940216
  30. XDS.
    Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):125-32 PMID: 20124692
  31. Mechanism of transfer RNA maturation by CCA-adding enzyme without using an oligonucleotide template.
    Nature. 2004 Aug 5;430(7000):640-5 PMID: 15295590
  32. RIG-I-dependent sensing of poly(dA:dT) through the induction of an RNA polymerase III-transcribed RNA intermediate.
    Nat Immunol. 2009 Oct;10(10):1065-72 PMID: 19609254
  33. Innate immune sensing of DNA viruses.
    Virology. 2011 Mar 15;411(2):153-62 PMID: 21334037
  34. STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling.
    Nature. 2008 Oct 2;455(7213):674-8 PMID: 18724357
  35. Phaser crystallographic software.
    J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674 PMID: 19461840
  36. The cytosolic nucleic acid sensor LRRFIP1 mediates the production of type I interferon via a beta-catenin-dependent pathway.
    Nat Immunol. 2010 Jun;11(6):487-94 PMID: 20453844
  37. STING is a direct innate immune sensor of cyclic di-GMP.
    Nature. 2011 Sep 25;478(7370):515-8 PMID: 21947006
  38. An orthogonal proteomic-genomic screen identifies AIM2 as a cytoplasmic DNA sensor for the inflammasome.
    Nat Immunol. 2009 Mar;10(3):266-72 PMID: 19158679
  39. RNA polymerase III detects cytosolic DNA and induces type I interferons through the RIG-I pathway.
    Cell. 2009 Aug 7;138(3):576-91 PMID: 19631370
Article Info
Journal
Nature
Abbr.
Nature
ISSN
1476-4687
DOI
10.1038/nature12305
Published
2013-06-20
Epub
2013-00-30
Pages
332-7
Language
English
Region
England
NLM ID
0410462
PMCID
PMC3768140
Subset
IM
Grants
European Research Council · 243046 · International
European Research Council · 322869 · International
NIAID NIH HHS · U19 AI083025 · United States
NIAID NIH HHS · U19AI083025 · United States
Databases
PDB
4JLX, 4JLZ, 4KB6
External Links
PubMed source DOI: 10.1038/nature12305 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2