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Viral SSB-bound ssDNA activates the bacterial anti-phage defense system DARNA

Puteikiene, R.; Vassallo, C.; Silanskas, A.; Juozapaitis, J.; Songailiene, I.; Laub, M. T.; Sasnauskas, G.

2026-02-23 microbiology
10.64898/2026.02.22.705581 bioRxiv
Show abstract

To protect themselves against phage infection, bacteria employ diverse defense systems that are typically activated specifically upon infection. However, the mechanisms of activation and self versus non-self discrimination for most systems remain poorly understood. Here, we show that the bacterial immunity protein DARNA, once activated, cleaves a subset of host tRNAs, thereby inhibiting phage propagation. Although phages escape DARNA-mediated defense through mutations in the gene encoding single-stranded DNA-binding protein (SSB), we find that phage SSBs do not directly stimulate DARNA. Instead, DARNA is activated by single-stranded DNA presented by phage SSB, but not by the host SSB. The recognition of an endogenous nucleic acid signal promoted by a viral protein ensures that DARNA can detect and respond to a broad range of viruses while avoiding auto-immunity.

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