Activation of Thoeris antiviral system via SIR2 effector filament assembly

Giedre Tamulaitiene; Dziugas Sabonis; Giedrius Sasnauskas; Audrone Ruksenaite; Arunas Silanskas; Carmel Avraham; Gal Ofir; Rotem Sorek; Mindaugas Zaremba; Virginijus Siksnys

doi:10.1038/s41586-024-07092-x

Activation of Thoeris antiviral system via SIR2 effector filament assembly

Giedre Tamulaitiene^*, Dziugas Sabonis, Giedrius Sasnauskas, Audrone Ruksenaite, Arunas Silanskas, Carmel Avraham, Gal Ofir, Rotem Sorek, Mindaugas Zaremba^*, Virginijus Siksnys^*

^*Corresponding author for this work

Department of Molecular Genetics

Research output: Contribution to journal › Article › peer-review

Abstract

To survive bacteriophage (phage) infections, bacteria developed numerous anti-phage defence systems^1–7. Some of them (for example, type III CRISPR–Cas, CBASS, Pycsar and Thoeris) consist of two modules: a sensor responsible for infection recognition and an effector that stops viral replication by destroying key cellular components^8–12. In the Thoeris system, a Toll/interleukin-1 receptor (TIR)-domain protein, ThsB, acts as a sensor that synthesizes an isomer of cyclic ADP ribose, 1′′−3′ glycocyclic ADP ribose (gcADPR), which is bound in the Smf/DprA-LOG (SLOG) domain of the ThsA effector and activates the silent information regulator 2 (SIR2)-domain-mediated hydrolysis of a key cell metabolite, NAD⁺ (refs. ^12–14). Although the structure of ThsA has been solved¹⁵, the ThsA activation mechanism remained incompletely understood. Here we show that 1′′−3′ gcADPR, synthesized in vitro by the dimeric ThsB′ protein, binds to the ThsA SLOG domain, thereby activating ThsA by triggering helical filament assembly of ThsA tetramers. The cryogenic electron microscopy (cryo-EM) structure of activated ThsA revealed that filament assembly stabilizes the active conformation of the ThsA SIR2 domain, enabling rapid NAD⁺ depletion. Furthermore, we demonstrate that filament formation enables a switch-like response of ThsA to the 1′′−3′ gcADPR signal.

Original language	English
Pages (from-to)	431-436
Number of pages	6
Journal	Nature
Volume	627
Issue number	8003
DOIs	https://doi.org/10.1038/s41586-024-07092-x
Publication status	Published Online - 21 Feb 2024

Bibliographical note

The research has been supported by the Research Council of Lithuania grant S-MIP-21-6 to G.T. The synchrotron X-ray diffraction data were collected at beamline P14 operated by the European Molecular Biology Laboratory, Hamburg at the PETRA III storage ring (Deutsches Elektronen-Synchrotron, Hamburg, Germany). Access to the beamline has been supported by iNEXT-Discovery, project number 871037, funded by the Horizon 2020 programme of the European Commission. We thank I. Drulyte and A. Carabias for discussions on cryo-EM structure determination.

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© The Author(s), under exclusive licence to Springer Nature Limited 2024.

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title = "Activation of Thoeris antiviral system via SIR2 effector filament assembly",

abstract = "To survive bacteriophage (phage) infections, bacteria developed numerous anti-phage defence systems1–7. Some of them (for example, type III CRISPR–Cas, CBASS, Pycsar and Thoeris) consist of two modules: a sensor responsible for infection recognition and an effector that stops viral replication by destroying key cellular components8–12. In the Thoeris system, a Toll/interleukin-1 receptor (TIR)-domain protein, ThsB, acts as a sensor that synthesizes an isomer of cyclic ADP ribose, 1′′−3′ glycocyclic ADP ribose (gcADPR), which is bound in the Smf/DprA-LOG (SLOG) domain of the ThsA effector and activates the silent information regulator 2 (SIR2)-domain-mediated hydrolysis of a key cell metabolite, NAD+ (refs. 12–14). Although the structure of ThsA has been solved15, the ThsA activation mechanism remained incompletely understood. Here we show that 1′′−3′ gcADPR, synthesized in vitro by the dimeric ThsB′ protein, binds to the ThsA SLOG domain, thereby activating ThsA by triggering helical filament assembly of ThsA tetramers. The cryogenic electron microscopy (cryo-EM) structure of activated ThsA revealed that filament assembly stabilizes the active conformation of the ThsA SIR2 domain, enabling rapid NAD+ depletion. Furthermore, we demonstrate that filament formation enables a switch-like response of ThsA to the 1′′−3′ gcADPR signal.",

author = "Giedre Tamulaitiene and Dziugas Sabonis and Giedrius Sasnauskas and Audrone Ruksenaite and Arunas Silanskas and Carmel Avraham and Gal Ofir and Rotem Sorek and Mindaugas Zaremba and Virginijus Siksnys",

note = "The research has been supported by the Research Council of Lithuania grant S-MIP-21-6 to G.T. The synchrotron X-ray diffraction data were collected at beamline P14 operated by the European Molecular Biology Laboratory, Hamburg at the PETRA III storage ring (Deutsches Elektronen-Synchrotron, Hamburg, Germany). Access to the beamline has been supported by iNEXT-Discovery, project number 871037, funded by the Horizon 2020 programme of the European Commission. We thank I. Drulyte and A. Carabias for discussions on cryo-EM structure determination. Publisher Copyright: {\textcopyright} The Author(s), under exclusive licence to Springer Nature Limited 2024.",

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doi = "10.1038/s41586-024-07092-x",

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AU - Tamulaitiene, Giedre

AU - Sabonis, Dziugas

AU - Sasnauskas, Giedrius

AU - Ruksenaite, Audrone

AU - Silanskas, Arunas

AU - Avraham, Carmel

AU - Ofir, Gal

AU - Sorek, Rotem

AU - Zaremba, Mindaugas

AU - Siksnys, Virginijus

N1 - The research has been supported by the Research Council of Lithuania grant S-MIP-21-6 to G.T. The synchrotron X-ray diffraction data were collected at beamline P14 operated by the European Molecular Biology Laboratory, Hamburg at the PETRA III storage ring (Deutsches Elektronen-Synchrotron, Hamburg, Germany). Access to the beamline has been supported by iNEXT-Discovery, project number 871037, funded by the Horizon 2020 programme of the European Commission. We thank I. Drulyte and A. Carabias for discussions on cryo-EM structure determination. Publisher Copyright: © The Author(s), under exclusive licence to Springer Nature Limited 2024.

PY - 2024/2/21

Y1 - 2024/2/21

N2 - To survive bacteriophage (phage) infections, bacteria developed numerous anti-phage defence systems1–7. Some of them (for example, type III CRISPR–Cas, CBASS, Pycsar and Thoeris) consist of two modules: a sensor responsible for infection recognition and an effector that stops viral replication by destroying key cellular components8–12. In the Thoeris system, a Toll/interleukin-1 receptor (TIR)-domain protein, ThsB, acts as a sensor that synthesizes an isomer of cyclic ADP ribose, 1′′−3′ glycocyclic ADP ribose (gcADPR), which is bound in the Smf/DprA-LOG (SLOG) domain of the ThsA effector and activates the silent information regulator 2 (SIR2)-domain-mediated hydrolysis of a key cell metabolite, NAD+ (refs. 12–14). Although the structure of ThsA has been solved15, the ThsA activation mechanism remained incompletely understood. Here we show that 1′′−3′ gcADPR, synthesized in vitro by the dimeric ThsB′ protein, binds to the ThsA SLOG domain, thereby activating ThsA by triggering helical filament assembly of ThsA tetramers. The cryogenic electron microscopy (cryo-EM) structure of activated ThsA revealed that filament assembly stabilizes the active conformation of the ThsA SIR2 domain, enabling rapid NAD+ depletion. Furthermore, we demonstrate that filament formation enables a switch-like response of ThsA to the 1′′−3′ gcADPR signal.

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Activation of Thoeris antiviral system via SIR2 effector filament assembly

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