Publications

Bacterial defense against phage predation involves diverse defense systems acting individually and concurrently, yet their interactions remain poorly understood. We investigated >100 defense systems in 42,925 bacterial genomes and identified numerous instances of their non-random co-occurrence and negative association. For several pairs of defense systems significantly co-occurring in Escherichia coli strains, we demonstrate synergistic anti-phage activity. Notably, Zorya II synergizes with Druantia III and ietAS defense systems, while tmn exhibits synergy with co-occurring systems Gabija, Septu I, and PrrC. For Gabija, tmn co-opts the sensory switch ATPase domain, enhancing anti-phage activity. Some defense system pairs that are negatively associated in E. coli show synergy and significantly co-occur in other taxa, demonstrating that bacterial immune repertoires are largely shaped by selection for resistance against host-specific phages rather than negative epistasis. Collectively, these findings demonstrate compatibility and synergy between defense systems, allowing bacteria to adopt flexible strategies for phage defense.

ABSTRACT Bacterial defence against phage predation involves diverse defence systems acting both individually and concurrently, yet their interactions remain poorly understood. We investigated >100 defence systems in 42,925 bacterial genomes and identified numerous instances of their non-random co-occurrence and mutual exclusion. For several pairs of defence systems significantly co-occurring in Escherichia coli strains, we demonstrate synergistic anti-phage activity, including tmn synergising with defence systems containing sensory switch ATPase domains. Some of the defence system pairs that are mutually exclusive in E. coli significantly co-occur in other taxa, and one tested mutually exclusive pair showed synergy. These findings demonstrate compatibility and synergy between defence systems, allowing bacteria to adopt flexible strategies for phage defence, driven by specific environmental pressures. The evolution of bacterial immune repertoires seems to be shaped largely by selection for resistance to host-specific phages rather than by negative epistasis.

Abstract Bacteria and archaea possess an enormous variety of antivirus immune systems that often share homologous proteins and domains, some of which contribute to diverse defense strategies. YprA family helicases are central to widespread defense systems DISARM, Dpd, and Druantia. Here, through comprehensive phylogenetic and structural prediction analysis of the YprA family, we identify several major, previously unrecognized clades, with unique signatures of domain architecture and associations with other genes. Each YprA family clade defines a distinct class of defense systems, which we denote ARMADA (dis ARM -related A ntiviral D efense A rray), BRIGADE ( B ase hypermodification and R estriction Involving G enes encoding A RMADA-like and D pd-like E ffectors), or TALON ( T OTE-like and A RMADA- L ike O peron with N uclease). In addition to the YprA-like helicase, ARMADA systems share two more proteins with DISARM. However, ARMADA YprA homologs are most similar to those of Druantia, suggesting ARMADA is a ‘missing link’ connecting DISARM and Druantia. We show experimentally that ARMADA protects bacteria against a broad range of phages via a direct, non-abortive mechanism. We also discovered multiple families of satellite phage-like mobile genetic elements that often carry both ARMADA and Druantia Type III systems and show that these can provide synergistic resistance against diverse phages.

Most bacteria encode multiple antiphage defence systems, but how these systems interact, remains poorly understood. Here, we define the mechanism of Druantia III and explore its synergy with Zorya II. Druantia III is a late-acting defence where DruH is the likely infection sensor and DruE is a helicase-nuclease effector that engages ssDNA-containing replication intermediates. Druantia III and Zorya II are each sensitive to the loss of RecD, whereas their synergy requires an intact RecBCD complex, indicating that the combined response depends on a shared DNA-processing hub. During T3 infection, Zorya apparently preserves this hub by limiting accumulation of the phage RecBCD-inhibitor Gp5.9, whereas DruE together with RecBCD promote the formation of DNA structures permissive for ZorE cleavage. During Bas37 infection, Druantia provides the dominant pathway and recruits ZorE in a non-canonical, ZorAB-independent manner. These findings show how shared DNA intermediates can connect defence systems into a coordinated antiphage response.

Abstract Membrane-associated phage defences remain poorly understood. Here we characterise Tmn, a YobI-family transmembrane P-loop NTPase that protects bacteria from phage infection by establishing a plasmolysis-associated antiviral state. Upon recognising phage T2 RIIB protein, Tmn enhances ATP turnover and selectively exports Mg 2+ , causing rapid cytoplasmic collapse that arrests phage replication without detectable membrane depolarisation or gross leakage of the cell content. Cryo-electron microscopy shows that Tmn assembles into a decameric membrane complex with extended cytosolic arms, an uncommon architecture among P-loop NTPases. The cytosolic arms, including a solenoid-like repeat domain, mediate trigger interaction and determine specificity. In addition to its primary defence function, Tmn-driven ATP collapse activates otherwise silent ATP-depletion-sensing defences, including Gabija and Septu type I, providing a mechanistic basis for synergy among defence systems and limiting secondary phage spread. These findings demonstrate that Tmn is a membrane-integrated sensor-effector that couples phage recognition to metabolic collapse and coordinated multi-layered immunity.

This article summarises the activities of the International Committee on Taxonomy of Viruses Bacterial Viruses Subcommittee, detailing developments in the classification of bacterial viruses. We provide here an overview of all new, abolished, moved and renamed taxa proposed in 2024, approved by the Executive Committee, and ratified by membership vote in 2025. Through the collective efforts of 74 international contributors of taxonomy proposals in this round, 43 ratified proposals have led to the creation of one new phylum, one class, four orders, 33 families, 14 subfamilies, 194 genera and 995 species. These proposals mark significant progress in refining the taxonomy of bacterial viruses. Key updates include the creation of new orders and families that include existing taxa to better reflect genomic and evolutionary relationships. As sequencing and bioinformatics approaches continue to advance, further expansion and refinements in viral taxonomy can be anticipated in the coming years.

Abstract The growing threat of multidrug-resistant Klebsiella pneumoniae, coupled with its role in gut colonisation, has intensified the search for new treatments, including bacteriophage therapy. Despite increasing documentation of Klebsiella-targeting phages, clinical applications remain limited, with key phage–bacteria interactions still poorly understood. A major obstacle is fragmented access to well-characterised phage–bacteria pairings, restricting the collective advancement of therapeutic and mechanistic insights. To address this gap, we created the Klebsiella Phage Collection (KlebPhaCol), an open resource comprising 52 phages and 74 Klebsiella isolates, characterised at phenotypic and genomic levels. These phages span six families—including a novel family, Felixviridae, associated with the human gut—and target 20 sequence types (including ST258, ST11, and ST14) and 19 capsular-locus types (including KL1 and KL2), across 6 Klebsiella species. Freely accessible at www.klebphacol.org, KlebPhaCol invites the scientific community to both use and contribute to this resource, fostering collaborative research and a deeper understanding of Klebsiella-phage interactions beyond therapeutic use.