A TIR-SAVED effector mediates antiviral immunity via a conserved host signal

Abstract

Cyclic oligonucleotide-based anti-phage signalling systems (CBASS) are widespread prokaryotic antiviral defense mechanisms that function through coordinated cyclase-effector interactions. Upon sensing viral infection, the cyclase produces a signal molecule that activates effector function and causes cell dormancy or death. However, the evolutionary origins and functional independence of CBASS components remain unclear. Type II CBASS systems commonly employ TIR-SAVED domain effector proteins that deplete cellular NAD+ during viral infection. Here, we demonstrate that a TIR-SAVED effector protein can operate as a standalone antiviral defense, causing significant growth inhibition and approximately 50% viral clearance during infection in the complete absence of its cognate cyclase. Remarkably, we show that the TIR-SAVED effector can sense cyclic di-AMP, a conserved second messenger produced by the host diadenylate cyclase DacZ, when the canonical CBASS signal is absent. This antiviral activity was associated with depletion of cellular NAD+ and required intact conserved functional residues within both the TIR and SAVED domains. These findings reveal a novel mechanism of antiviral signalling that expands the functional repertoire of CBASS. They also provide insights into the modular evolution of complex prokaryotic immune systems, suggesting that what are now CBASS effectors might have evolved as independent defense components before being integrated into multi-protein systems.