Catalytic Promiscuity of CGAS: A Facile Enzymatic Synthesis of 2'-3'-Linked Cyclic Dinucleotides

Overview

Journal Chembiochem

Specialty Biochemistry

Date 2020 Jul 8

PMID 32633874

Citations 10

Authors

Katrin Rosenthal

Martin Becker

Jascha Rolf

Regine Siedentop

Michael Hillen

Markus Nett

Stephan Lutz

Affiliations

Soon will be listed here.

Abstract

Cyclic GMP-AMP synthase (cGAS) is a cytosolic DNA sensor that catalyzes the synthesis of the cyclic GMP-AMP dinucleotide 2'3'-cGAMP. 2'3'-cGAMP functions as inducer for the production of type I interferons. Derivatives of this important second messenger are highly valuable for pharmaceutical applications. However, the production of these analogues requires complex, multistep syntheses. Herein, human cGAS is shown to react with a series of unnatural nucleotides, thus leading to novel cyclic dinucleotides. Most substrate derivatives with modifications at the nucleobase, ribose, and the α-thio phosphate were accepted. These results demonstrate the catalytic promiscuity of human cGAS and its utility for the biocatalytic synthesis of cyclic dinucleotide derivatives.

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References

Kato K, Ishii R, Goto E, Ishitani R, Tokunaga F, Nureki O . Structural and functional analyses of DNA-sensing and immune activation by human cGAS. PLoS One. 2013; 8(10):e76983. PMC: 3792152. DOI: 10.1371/journal.pone.0076983. View

Veliath E, Kim S, Gaffney B, Jones R . Synthesis and characterization of C8 analogs of c-di-GMP. Nucleosides Nucleotides Nucleic Acids. 2011; 30(11):961-78. DOI: 10.1080/15257770.2011.624567. View

Danilchanka O, Mekalanos J . Cyclic dinucleotides and the innate immune response. Cell. 2013; 154(5):962-970. PMC: 3931520. DOI: 10.1016/j.cell.2013.08.014. View

Civril F, Deimling T, de Oliveira Mann C, Ablasser A, Moldt M, Witte G . Structural mechanism of cytosolic DNA sensing by cGAS. Nature. 2013; 498(7454):332-7. PMC: 3768140. DOI: 10.1038/nature12305. View

Pettersen E, Goddard T, Huang C, Couch G, Greenblatt D, Meng E . UCSF Chimera--a visualization system for exploratory research and analysis. J Comput Chem. 2004; 25(13):1605-12. DOI: 10.1002/jcc.20084. View

Gaffney B, Stebbins N, Jones R . Synthesis of biotinylated c-di-gmp and c-di-amp using click conjugation. Nucleosides Nucleotides Nucleic Acids. 2013; 32(1):1-16. PMC: 3561633. DOI: 10.1080/15257770.2012.748196. View

Khersonsky O, Tawfik D . Enzyme promiscuity: a mechanistic and evolutionary perspective. Annu Rev Biochem. 2010; 79:471-505. DOI: 10.1146/annurev-biochem-030409-143718. View

Gaffney B, Veliath E, Zhao J, Jones R . One-flask syntheses of c-di-GMP and the [Rp,Rp] and [Rp,Sp] thiophosphate analogues. Org Lett. 2010; 12(14):3269-71. PMC: 2905038. DOI: 10.1021/ol101236b. View

Gao P, Ascano M, Wu Y, Barchet W, Gaffney B, Zillinger T . Cyclic [G(2',5')pA(3',5')p] is the metazoan second messenger produced by DNA-activated cyclic GMP-AMP synthase. Cell. 2013; 153(5):1094-107. PMC: 4382009. DOI: 10.1016/j.cell.2013.04.046. View

10.

Ablasser A, Chen Z . cGAS in action: Expanding roles in immunity and inflammation. Science. 2019; 363(6431). DOI: 10.1126/science.aat8657. View

11.

Pandya C, Farelli J, Dunaway-Mariano D, Allen K . Enzyme promiscuity: engine of evolutionary innovation. J Biol Chem. 2014; 289(44):30229-30236. PMC: 4215207. DOI: 10.1074/jbc.R114.572990. View

12.

Zhang X, Shi H, Wu J, Zhang X, Sun L, Chen C . Cyclic GMP-AMP containing mixed phosphodiester linkages is an endogenous high-affinity ligand for STING. Mol Cell. 2013; 51(2):226-35. PMC: 3808999. DOI: 10.1016/j.molcel.2013.05.022. View

13.

Xia P, Ye B, Wang S, Zhu X, Du Y, Xiong Z . Glutamylation of the DNA sensor cGAS regulates its binding and synthase activity in antiviral immunity. Nat Immunol. 2016; 17(4):369-78. DOI: 10.1038/ni.3356. View

14.

Kranzusch P, Lee A, Wilson S, Solovykh M, Vance R, Berger J . Structure-guided reprogramming of human cGAS dinucleotide linkage specificity. Cell. 2014; 158(5):1011-1021. PMC: 4157622. DOI: 10.1016/j.cell.2014.07.028. View

15.

Watson R, Bell S, MacDuff D, Kimmey J, Diner E, Olivas J . The Cytosolic Sensor cGAS Detects Mycobacterium tuberculosis DNA to Induce Type I Interferons and Activate Autophagy. Cell Host Microbe. 2015; 17(6):811-819. PMC: 4466081. DOI: 10.1016/j.chom.2015.05.004. View

16.

Bornscheuer U, Kazlauskas R . Catalytic promiscuity in biocatalysis: using old enzymes to form new bonds and follow new pathways. Angew Chem Int Ed Engl. 2004; 43(45):6032-40. DOI: 10.1002/anie.200460416. View

17.

Gaffney B, Jones R . Synthesis of c-di-GMP analogs with thiourea, urea, carbodiimide, and guanidinium linkages. Org Lett. 2013; 16(1):158-61. PMC: 3933394. DOI: 10.1021/ol403154w. View

18.

Hall J, Ralph E, Shanker S, Wang H, Byrnes L, Horst R . The catalytic mechanism of cyclic GMP-AMP synthase (cGAS) and implications for innate immunity and inhibition. Protein Sci. 2017; 26(12):2367-2380. PMC: 5699495. DOI: 10.1002/pro.3304. View

19.

Zhang X, Wu J, Du F, Xu H, Sun L, Chen Z . The cytosolic DNA sensor cGAS forms an oligomeric complex with DNA and undergoes switch-like conformational changes in the activation loop. Cell Rep. 2014; 6(3):421-30. PMC: 3969844. DOI: 10.1016/j.celrep.2014.01.003. View

20.

Rosenthal K, Becker M, Rolf J, Siedentop R, Hillen M, Nett M . Catalytic Promiscuity of cGAS: A Facile Enzymatic Synthesis of 2'-3'-Linked Cyclic Dinucleotides. Chembiochem. 2020; 21(22):3225-3228. PMC: 7754487. DOI: 10.1002/cbic.202000433. View