Modularity of Zorya defense systems during phage inhibition

Mariano, Giuseppina; Deme, Justin C.; Readshaw, Jennifer J.; Grobbelaar, Matthew J.; Keenan, Mackenzie; El-Masri, Yasmin; Bamford, Lindsay; Songra, Suraj; Blower, Tim R.; Palmer, Tracy; Lea, Susan M.

Abstract

Bacteria have evolved an extraordinary diversity of defense systems against bacteriophage (phage) predation. However, the molecular mechanisms underlying these anti-phage systems often remain elusive. Here, we provide mechanistic and structural insights into Zorya phage defense systems. Using cryo-EM structural analyses, we show that the Zorya type I and II core components, ZorA and ZorB, assemble in a 5:2 complex that is similar to inner-membrane ion-driven, rotary motors that power flagellar rotation, type 9 secretion, gliding and the Ton nutrient uptake systems. The ZorAB complex has an elongated cytoplasmic tail assembled by bundling the C-termini of the five ZorA subunits. Mutagenesis demonstrates that peptidoglycan binding by the periplasmic domains of ZorB, the structured cytoplasmic tail of ZorA, and ion flow through the motor is important for function in both type I and II systems. Furthermore, we identify ZorE as the effector module of the Zorya II system, possessing nickase activity. Our work reveals the molecular basis of the activity of Zorya systems and highlights the ZorE nickase as crucial for population-wide immunity in the type II system.

Citation

Mariano, G., Deme, J. C., Readshaw, J. J., Grobbelaar, M. J., Keenan, M., El-Masri, Y., Bamford, L., Songra, S., Blower, T. R., Palmer, T., & Lea, S. M. (2025). Modularity of Zorya defense systems during phage inhibition. Nature Communications, 16(1), Article 2344. https://doi.org/10.1038/s41467-025-57397-2

• Journal Article Type: Article
• Acceptance Date: Feb 17, 2025
• Online Publication Date: Mar 8, 2025
• Publication Date: Mar 8, 2025
• Deposit Date: Mar 17, 2025
• Publicly Available Date: Mar 17, 2025
• Journal: Nature Communications
• Electronic ISSN: 2041-1723
• Publisher: Nature Research
• Peer Reviewed: Peer Reviewed
• Volume: 16
• Issue: 1
• Article Number: 2344
• DOI: https://doi.org/10.1038/s41467-025-57397-2
• Public URL: https://durham-repository.worktribe.com/output/3714608