cGAS-like receptors sense RNA and control 3'2'-cGAMP signalling in Drosophila

Slavik, Kailey M.; Morehouse, Benjamin R.; Ragucci, Adelyn E.; Zhou, Wen; Ai, Xianlong; Chen, Yuqiang; Li, Lihua; Wei, Ziming; Bähre, Heike; König, Martin; Seifert, Roland; Lee, Amy S. Y.; Cai, Hua; Imler, Jean-Luc; Kranzusch, Philip J.

doi:10.1038/s41586-021-03743-5

cGAS-like receptors sense RNA and control 3'2'-cGAMP signalling in Drosophila

Journal Article · Wed Jul 14 00:00:00 EDT 2021 · Nature (London)

DOI:https://doi.org/10.1038/s41586-021-03743-5· OSTI ID:1820446

Slavik, Kailey M. ^[1]; ^[1]; Ragucci, Adelyn E. ^[1]; Zhou, Wen ^[1]; Ai, Xianlong ^[2]; Chen, Yuqiang ^[2]; Li, Lihua ^[2]; Wei, Ziming ^[2]; Bähre, Heike ^[3]; König, Martin ^[3]; Seifert, Roland ^[3]; Lee, Amy S. Y. ^[4]; Cai, Hua ^[2]; Imler, Jean-Luc ^[5]; ^[1]

Harvard Medical School, Boston, MA (United States); Dana-Farber Cancer Institute, Boston, MA (United States)
Guangzhou Medical University (China)
Hannover Medical School (Germany)
Dana-Farber Cancer Institute, Boston, MA (United States); Harvard Medical School, Boston, MA (United States)
Guangzhou Medical University (China); Université de Strasbourg (France)

Cyclic GMP–AMP synthase (cGAS) is a cytosolic DNA sensor that produces the second messenger cG[2'–5']pA[3'–5']p (2'3'-cGAMP) and controls activation of innate immunity in mammalian cells. Animal genomes typically encode multiple proteins with predicted homology to cGAS, but the function of these uncharacterized enzymes is unknown. Here we show that cGAS-like receptors (cGLRs) are innate immune sensors that are capable of recognizing divergent molecular patterns and catalysing synthesis of distinct nucleotide second messenger signals. Crystal structures of human and insect cGLRs reveal a nucleotidyltransferase signalling core shared with cGAS and a diversified primary ligand-binding surface modified with notable insertions and deletions. We demonstrate that surface remodelling of cGLRs enables altered ligand specificity and used a forward biochemical screen to identify cGLR1 as a double-stranded RNA sensor in the model organism Drosophila melanogaster. We show that RNA recognition activates Drosophila cGLR1 to synthesize the novel product cG[3'–5']pA[2'–5']p (3'2'-cGAMP). A crystal structure of Drosophila stimulator of interferon genes (dSTING) in complex with 3'2'-cGAMP explains selective isomer recognition, and 3'2'-cGAMP induces an enhanced antiviral state in vivo that protects from viral infection. Similar to radiation of Toll-like receptors in pathogen immunity, our results establish cGLRs as a diverse family of metazoan pattern recognition receptors.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: AC02-06CH11357; AC02-05CH11231

OSTI ID:: 1820446

Journal Information:: Nature (London), Journal Name: Nature (London) Journal Issue: 7874 Vol. 597; ISSN 0028-0836

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

References (51)

Overexpression of wild type or a Q311E mutant MB21D2 promotes a pro‐oncogenic phenotype in HNSCC Gracilla, Daniel E.; Korla, Praveen Kumar; Lai, Ming‐Tsung Molecular Oncology, Vol. 14, Issue 12 https://doi.org/10.1002/1878-0261.12806	journal	October 2020
Protein Sequence Analysis Using the MPI Bioinformatics Toolkit Gabler, Felix; Nam, Seung‐Zin; Till, Sebastian Current Protocols in Bioinformatics, Vol. 72, Issue 1 https://doi.org/10.1002/cpbi.108	journal	December 2020
DALI and the persistence of protein shape Holm, Liisa Protein Science, Vol. 29, Issue 1 https://doi.org/10.1002/pro.3749	journal	November 2019
PROMALS3D: Multiple Protein Sequence Alignment Enhanced with Evolutionary and Three-Dimensional Structural Information Pei, Jimin; Grishin, Nick V. Methods in Molecular Biology https://doi.org/10.1007/978-1-62703-646-7_17	book	August 2013
Cyclic [G(2′,5′)pA(3′,5′)p] Is the Metazoan Second Messenger Produced by DNA-Activated Cyclic GMP-AMP Synthase Gao, Pu; Ascano, Manuel; Wu, Yang Cell, Vol. 153, Issue 5 https://doi.org/10.1016/j.cell.2013.04.046	journal	May 2013
Structure-Guided Reprogramming of Human cGAS Dinucleotide Linkage Specificity Kranzusch, Philip J.; Lee, Amy S. Y.; Wilson, Stephen C. Cell, Vol. 158, Issue 5 https://doi.org/10.1016/j.cell.2014.07.028	journal	August 2014
Structure of the Human cGAS–DNA Complex Reveals Enhanced Control of Immune Surveillance Zhou, Wen; Whiteley, Aaron T.; de Oliveira Mann, Carina C. Cell, Vol. 174, Issue 2 https://doi.org/10.1016/j.cell.2018.06.026	journal	July 2018
CBASS Immunity Uses CARF-Related Effectors to Sense 3′–5′- and 2′–5′-Linked Cyclic Oligonucleotide Signals and Protect Bacteria from Phage Infection Lowey, Brianna; Whiteley, Aaron T.; Keszei, Alexander F. A. Cell, Vol. 182, Issue 1 https://doi.org/10.1016/j.cell.2020.05.019	journal	July 2020
Structure of Human cGAS Reveals a Conserved Family of Second-Messenger Enzymes in Innate Immunity Kranzusch, Philip J.; Lee, Amy Si-Ying; Berger, James M. Cell Reports, Vol. 3, Issue 5 https://doi.org/10.1016/j.celrep.2013.05.008	journal	May 2013
The Innate Immune DNA Sensor cGAS Produces a Noncanonical Cyclic Dinucleotide that Activates Human STING Diner, Elie J.; Burdette, Dara L.; Wilson, Stephen C. Cell Reports, Vol. 3, Issue 5 https://doi.org/10.1016/j.celrep.2013.05.009	journal	May 2013
The Cytosolic DNA Sensor cGAS Forms an Oligomeric Complex with DNA and Undergoes Switch-like Conformational Changes in the Activation Loop Zhang, Xu; Wu, Jiaxi; Du, Fenghe Cell Reports, Vol. 6, Issue 3 https://doi.org/10.1016/j.celrep.2014.01.003	journal	February 2014
Analysis of Drosophila STING Reveals an Evolutionarily Conserved Antimicrobial Function Martin, Marina; Hiroyasu, Aoi; Guzman, R. Marena Cell Reports, Vol. 23, Issue 12 https://doi.org/10.1016/j.celrep.2018.05.029	journal	June 2018
Mn2+ Directly Activates cGAS and Structural Analysis Suggests Mn2+ Induces a Noncanonical Catalytic Synthesis of 2′3′-cGAMP Zhao, Zhen; Ma, Zhixing; Wang, Bo Cell Reports, Vol. 32, Issue 7 https://doi.org/10.1016/j.celrep.2020.108053	journal	August 2020
Molecular basis of CD-NTase nucleotide selection in CBASS anti-phage defense Govande, Apurva A.; Duncan-Lowey, Brianna; Eaglesham, James B. Cell Reports, Vol. 35, Issue 9 https://doi.org/10.1016/j.celrep.2021.109206	journal	June 2021
Inflammation-Induced, STING-Dependent Autophagy Restricts Zika Virus Infection in the Drosophila Brain Liu, Yuan; Gordesky-Gold, Beth; Leney-Greene, Michael Cell Host & Microbe, Vol. 24, Issue 1 https://doi.org/10.1016/j.chom.2018.05.022	journal	July 2018
Sensing and signalling viral infection in drosophila Schneider, Juliette; Imler, Jean-Luc Developmental & Comparative Immunology, Vol. 117 https://doi.org/10.1016/j.dci.2020.103985	journal	April 2021
The Kinase IKKβ Regulates a STING- and NF-κB-Dependent Antiviral Response Pathway in Drosophila Goto, Akira; Okado, Kiyoshi; Martins, Nelson Immunity, Vol. 49, Issue 2 https://doi.org/10.1016/j.immuni.2018.07.013	journal	August 2018
Cyclic GMP-AMP Containing Mixed Phosphodiester Linkages Is An Endogenous High-Affinity Ligand for STING Zhang, Xu; Shi, Heping; Wu, Jiaxi Molecular Cell, Vol. 51, Issue 2 https://doi.org/10.1016/j.molcel.2013.05.022	journal	July 2013
Ancient Origin of cGAS-STING Reveals Mechanism of Universal 2′,3′ cGAMP Signaling Kranzusch, Philip J.; Wilson, Stephen C.; Lee, Amy S. Y. Molecular Cell, Vol. 59, Issue 6 https://doi.org/10.1016/j.molcel.2015.07.022	journal	September 2015
HORMA Domain Proteins and a Trip13-like ATPase Regulate Bacterial cGAS-like Enzymes to Mediate Bacteriophage Immunity Ye, Qiaozhen; Lau, Rebecca K.; Mathews, Ian T. Molecular Cell, Vol. 77, Issue 4 https://doi.org/10.1016/j.molcel.2019.12.009	journal	February 2020
cGAS phase separation inhibits TREX1-mediated DNA degradation and enhances cytosolic DNA sensing Zhou, Wen; Mohr, Lisa; Maciejowski, John Molecular Cell, Vol. 81, Issue 4 https://doi.org/10.1016/j.molcel.2021.01.024	journal	February 2021
cGAS and CD-NTase enzymes: structure, mechanism, and evolution Kranzusch, Philip J. Current Opinion in Structural Biology, Vol. 59 https://doi.org/10.1016/j.sbi.2019.08.003	journal	December 2019
The Activation Mechanism of 2′-5′-Oligoadenylate Synthetase Gives New Insights Into OAS/cGAS Triggers of Innate Immunity Lohöfener, Jan; Steinke, Nicola; Kay-Fedorov, Penelope Structure, Vol. 23, Issue 5 https://doi.org/10.1016/j.str.2015.03.012	journal	May 2015
Structural mechanism of cytosolic DNA sensing by cGAS Civril, Filiz; Deimling, Tobias; de Oliveira Mann, Carina C. Nature, Vol. 498, Issue 7454 https://doi.org/10.1038/nature12305	journal	May 2013
cGAS produces a 2′-5′-linked cyclic dinucleotide second messenger that activates STING Ablasser, Andrea; Goldeck, Marion; Cavlar, Taner Nature, Vol. 498, Issue 7454 https://doi.org/10.1038/nature12306	journal	May 2013
Distinct patterns of somatic genome alterations in lung adenocarcinomas and squamous cell carcinomas Campbell, Joshua D.; Alexandrov, Anton; Kim, Jaegil Nature Genetics, Vol. 48, Issue 6 https://doi.org/10.1038/ng.3564	journal	May 2016
Fiji: an open-source platform for biological-image analysis Schindelin, Johannes; Arganda-Carreras, Ignacio; Frise, Erwin Nature Methods, Vol. 9, Issue 7 https://doi.org/10.1038/nmeth.2019	journal	June 2012
The Phyre2 web portal for protein modeling, prediction and analysis Kelley, Lawrence A.; Mezulis, Stefans; Yates, Christopher M. Nature Protocols, Vol. 10, Issue 6 https://doi.org/10.1038/nprot.2015.053	journal	May 2015
Small RNA-based antimicrobial immunity Guo, Zhongxin; Li, Yang; Ding, Shou-Wei Nature Reviews Immunology, Vol. 19, Issue 1 https://doi.org/10.1038/s41577-018-0071-x	journal	October 2018
Viral and metazoan poxins are cGAMP-specific nucleases that restrict cGAS–STING signalling Eaglesham, James B.; Pan, Youdong; Kupper, Thomas S. Nature, Vol. 566, Issue 7743 https://doi.org/10.1038/s41586-019-0928-6	journal	February 2019
Bacterial cGAS-like enzymes synthesize diverse nucleotide signals Whiteley, Aaron T.; Eaglesham, James B.; de Oliveira Mann, Carina C. Nature, Vol. 567, Issue 7747 https://doi.org/10.1038/s41586-019-0953-5	journal	February 2019
Autophagy induction via STING trafficking is a primordial function of the cGAS pathway Gui, Xiang; Yang, Hui; Li, Tuo Nature, Vol. 567, Issue 7747 https://doi.org/10.1038/s41586-019-1006-9	journal	March 2019
Cyclic GMP–AMP signalling protects bacteria against viral infection Cohen, Daniel; Melamed, Sarah; Millman, Adi Nature, Vol. 574, Issue 7780 https://doi.org/10.1038/s41586-019-1605-5	journal	September 2019
STING cyclic dinucleotide sensing originated in bacteria Morehouse, Benjamin R.; Govande, Apurva A.; Millman, Adi Nature, Vol. 586, Issue 7829 https://doi.org/10.1038/s41586-020-2719-5	journal	September 2020
Two cGAS-like receptors induce antiviral immunity in Drosophila Holleufer, Andreas; Winther, Kasper Grønbjerg; Gad, Hans Henrik Nature, Vol. 597, Issue 7874 https://doi.org/10.1038/s41586-021-03800-z	journal	July 2021
Structural and biochemical characterization of the cell fate determining nucleotidyltransferase fold protein MAB21L1 de Oliveira Mann, Carina C.; Kiefersauer, Reiner; Witte, Gregor Scientific Reports, Vol. 6, Issue 1 https://doi.org/10.1038/srep27498	journal	June 2016
Structural basis for cytosolic double-stranded RNA surveillance by human oligoadenylate synthetase 1 Donovan, J.; Dufner, M.; Korennykh, A. Proceedings of the National Academy of Sciences, Vol. 110, Issue 5 https://doi.org/10.1073/pnas.1218528110	journal	January 2013
Human cGAS catalytic domain has an additional DNA-binding interface that enhances enzymatic activity and liquid-phase condensation Xie, Wei; Lama, Lodoe; Adura, Carolina Proceedings of the National Academy of Sciences https://doi.org/10.1073/pnas.1905013116	journal	May 2019
MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability Katoh, K.; Standley, D. M. Molecular Biology and Evolution, Vol. 30, Issue 4 https://doi.org/10.1093/molbev/mst010	journal	January 2013
Molecular evolutionary and structural analysis of the cytosolic DNA sensor cGAS and STING Wu, Xiaomei; Wu, Fei-Hua; Wang, Xiaoqiang Nucleic Acids Research, Vol. 42, Issue 13 https://doi.org/10.1093/nar/gku569	journal	June 2014
Comparative genomic analyses reveal a vast, novel network of nucleotide-centric systems in biological conflicts, immunity and signaling Burroughs, A. Maxwell; Zhang, Dapeng; Schäffer, Daniel E. Nucleic Acids Research, Vol. 43, Issue 22 https://doi.org/10.1093/nar/gkv1267	journal	November 2015
Coot model-building tools for molecular graphics Emsley, Paul; Cowtan, Kevin Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12, p. 2126-2132 https://doi.org/10.1107/S0907444904019158	journal	November 2004
XDS Kabsch, Wolfgang Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 2 https://doi.org/10.1107/S0907444909047337	journal	January 2010
Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix Liebschner, Dorothee; Afonine, Pavel V.; Baker, Matthew L. Acta Crystallographica Section D Structural Biology, Vol. 75, Issue 10 https://doi.org/10.1107/S2059798319011471	journal	October 2019
c-di-AMP Secreted by Intracellular Listeria monocytogenes Activates a Host Type I Interferon Response Woodward, J. J.; Iavarone, A. T.; Portnoy, D. A. Science, Vol. 328, Issue 5986 https://doi.org/10.1126/science.1189801	journal	May 2010
Cyclic GMP-AMP Synthase Is a Cytosolic DNA Sensor That Activates the Type I Interferon Pathway Sun, L.; Wu, J.; Du, F. Science, Vol. 339, Issue 6121 https://doi.org/10.1126/science.1232458	journal	December 2012
DNA-induced liquid phase condensation of cGAS activates innate immune signaling Du, Mingjian; Chen, Zhijian J. Science, Vol. 361, Issue 6403 https://doi.org/10.1126/science.aat1022	journal	July 2018
2′3′-cGAMP triggers a STING- and NF-κB–dependent broad antiviral response in Drosophila Cai, Hua; Holleufer, Andreas; Simonsen, Bine Science Signaling, Vol. 13, Issue 660 https://doi.org/10.1126/scisignal.abc4537	journal	December 2020
Innate Immune Sensing and Signaling of Cytosolic Nucleic Acids Wu, Jiaxi; Chen, Zhijian J. Annual Review of Immunology, Vol. 32, Issue 1 https://doi.org/10.1146/annurev-immunol-032713-120156	journal	March 2014
The Toll-Dorsal Pathway Is Required for Resistance to Viral Oral Infection in Drosophila Ferreira, Álvaro Gil; Naylor, Huw; Esteves, Sara Santana PLoS Pathogens, Vol. 10, Issue 12 https://doi.org/10.1371/journal.ppat.1004507	journal	December 2014
Structures of diverse poxin cGAMP nucleases reveal a widespread role for cGAS-STING evasion in host–pathogen conflict Eaglesham, James B.; McCarty, Kacie L.; Kranzusch, Philip J. eLife, Vol. 9 https://doi.org/10.7554/eLife.59753	journal	November 2020

Similar Records

Structures of diverse poxin cGAMP nucleases reveal a widespread role for cGAS-STING evasion in host–pathogen conflict

Journal Article · Sun Nov 15 23:00:00 EST 2020 · eLife · OSTI ID:1736294

Viral and metazoan poxins are cGAMP-specific nucleases that restrict cGAS-STING signalling

Journal Article · Tue Feb 05 23:00:00 EST 2019 · Nature (London) · OSTI ID:1497105

Structural Basis for Molecular Discrimination by a 3',3'-cGAMP Sensing Riboswitch

Journal Article · Tue Apr 07 00:00:00 EDT 2015 · Cell Reports · OSTI ID:1178128

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
innate immunity
structural biology

cGAS-like receptors sense RNA and control 3'2'-cGAMP signalling in Drosophila

Citation Formats

References (51)

Similar Records

Related Subjects