Ancient Origin of cGAS-STING Reveals Mechanism of Universal 2′,3′ cGAMP Signaling

In humans, the cGAS-STING immunity pathway signals in response to cytosolic DNA via 2′,3′ cGAMP, a cyclic dinucleotide (CDN) second messenger containing mixed 2′–5′ and 3′–5′ phosphodiester bonds. Prokaryotes also produce CDNs, but these are exclusively 3′ linked, and thus the evolutionary origins of human 2′,3′ cGAMP signaling are unknown. Here we illuminate the ancient origins of human cGAMP signaling by discovery of a functional cGAS-STING pathway in Nematostella vectensis, an anemone species >500 million years diverged from humans. Anemone cGAS appears to produce a 3′,3′ CDN that anemone STING recognizes through nucleobase-specific contacts not observed in human STING. Nevertheless, anemone STING binds mixed-linkage 2′,3′ cGAMP indistinguishably from human STING, trapping a unique structural conformation not induced by 3′,3′ CDNs. These results reveal that human mixed-linkage cGAMP achieves universal signaling by exploiting a deeply conserved STING conformational intermediate, providing critical insight for therapeutic targeting of the STING pathway. [Display omitted] •Binding of CDNs is an evolutionarily ancient STING function, predating interferons•cGAS-STING function is conserved in anemone, >500 million years diverged from humans•Anemone cGAS produces a canonical 3′,3′ linked CDN similar to those in bacteria•Vertebrate 2′,3′ cGAMP signaling exploits a deeply conserved STING conformation Kranzusch and Wilson et al. use structural and biochemical approaches to characterize human and anemone cGAS-STING immune pathways, demonstrating that the product of human cGAS is a potent STING activator because it targets an ancient, conserved, intermediate conformation of STING.