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bioRxiv · 10.1101/2024.02.11.579819

Phages reconstitute NAD+ to counter bacterial immunity

Osterman, I.·Samra, H.·Rousset, F.·Loseva, E.·Itkin, M.·Malitsky, S.·Yirmiya, E.·Millman, A.·Sorek, R.
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Abstract

Bacteria defend against phage infection via a variety of antiphage defense systems. Many defense systems were recently shown to deplete cellular nicotinamide adenine dinucleotide (NAD+) in response to infection, by breaking NAD+ to ADP-ribose (ADPR) and nicotinamide. It was demonstrated that NAD+ depletion during infection deprives the phage from this essential molecule and impedes phage replication. Here we show that a substantial fraction of phages possess enzymatic pathways allowing reconstitution of NAD+ from its degradation products in infected cells. We describe NAD+ reconstitution pathway 1 (NARP1), a two-step pathway in which one enzyme phosphorylates ADPR to generate ADPR-pyrophosphate (ADPR-PP), and the second enzyme conjugates ADPR- PP and nicotinamide to generate NAD+. Phages encoding the NARP1 pathway can overcome a diverse set of defense systems, including Thoeris, DSR1, DSR2, SIR2-HerA, and SEFIR, all of which deplete NAD+ as part of their defensive mechanism. Phylogenetic analyses show that NARP1 is primarily encoded on phage genomes, suggesting a phage- specific function in countering bacterial defenses. A second pathway, NARP2, allows phages to overcome bacterial defenses by building NAD+ via metabolites different than ADPR-PP. Our findings report a unique immune evasion strategy where viruses rebuild molecules depleted by defense systems, thus overcoming host immunity.

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Osterman, I., Samra, H., Rousset, F., Loseva, E., Itkin, M., Malitsky, S., Yirmiya, E., Millman, A., Sorek, R.. 2024-02-13. Phages reconstitute NAD+ to counter bacterial immunity. https://doi.org/10.1101/2024.02.11.579819

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