Cryo-EM structure of the RADAR supramolecular anti-phage defense complex

Brianna Duncan-Lowey; Nitzan Tal; Alex G. Johnson; Shaun Rawson; Megan L. Mayer; Shany Doron; Adi Millman; Sarah Melamed; Taya Fedorenko; Assaf Kacen; Alexander Brandis; Tevie Mehlman; Gil Amitai; Rotem Sorek; Philip J. Kranzusch

doi:10.1016/j.cell.2023.01.012

Cryo-EM structure of the RADAR supramolecular anti-phage defense complex

Brianna Duncan-Lowey, Nitzan Tal, Alex G. Johnson, Shaun Rawson, Megan L. Mayer, Shany Doron, Adi Millman, Sarah Melamed, Taya Fedorenko, Assaf Kacen, Alexander Brandis, Tevie Mehlman, Gil Amitai, Rotem Sorek, Philip J. Kranzusch

Weizmann Institute of Science

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

Abstract

RADAR is a two-protein bacterial defense system that was reported to defend against phage by “editing” messenger RNA. Here, we determine cryo-EM structures of the RADAR defense complex, revealing RdrA as a heptameric, two-layered AAA+ ATPase and RdrB as a dodecameric, hollow complex with twelve surface-exposed deaminase active sites. RdrA and RdrB join to form a giant assembly up to 10 MDa, with RdrA docked as a funnel over the RdrB active site. Surprisingly, our structures reveal an RdrB active site that targets mononucleotides. We show that RdrB catalyzes ATP-to-ITP conversion in vitro and induces the massive accumulation of inosine mononucleotides during phage infection in vivo, limiting phage replication. Our results define ATP mononucleotide deamination as a determinant of RADAR immunity and reveal supramolecular assembly of a nucleotide-modifying machine as a mechanism of anti-phage defense.

Original language	English
Pages (from-to)	987-998.e15
Journal	Cell
Volume	186
Issue number	5
Early online date	9 Feb 2023
DOIs	https://doi.org/10.1016/j.cell.2023.01.012
State	Published - 2 Mar 2023

All Science Journal Classification (ASJC) codes

General Biochemistry,Genetics and Molecular Biology

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rs_Cell_CryoEMStructureoftheRADAR_PV2023Final published version, 9.4 MBLicense: CC BY

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title = "Cryo-EM structure of the RADAR supramolecular anti-phage defense complex",

abstract = "RADAR is a two-protein bacterial defense system that was reported to defend against phage by “editing” messenger RNA. Here, we determine cryo-EM structures of the RADAR defense complex, revealing RdrA as a heptameric, two-layered AAA+ ATPase and RdrB as a dodecameric, hollow complex with twelve surface-exposed deaminase active sites. RdrA and RdrB join to form a giant assembly up to 10 MDa, with RdrA docked as a funnel over the RdrB active site. Surprisingly, our structures reveal an RdrB active site that targets mononucleotides. We show that RdrB catalyzes ATP-to-ITP conversion in vitro and induces the massive accumulation of inosine mononucleotides during phage infection in vivo, limiting phage replication. Our results define ATP mononucleotide deamination as a determinant of RADAR immunity and reveal supramolecular assembly of a nucleotide-modifying machine as a mechanism of anti-phage defense.",

author = "Brianna Duncan-Lowey and Nitzan Tal and Johnson, {Alex G.} and Shaun Rawson and Mayer, {Megan L.} and Shany Doron and Adi Millman and Sarah Melamed and Taya Fedorenko and Assaf Kacen and Alexander Brandis and Tevie Mehlman and Gil Amitai and Rotem Sorek and Kranzusch, {Philip J.}",

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doi = "10.1016/j.cell.2023.01.012",

language = "الإنجليزيّة",

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T1 - Cryo-EM structure of the RADAR supramolecular anti-phage defense complex

AU - Duncan-Lowey, Brianna

AU - Tal, Nitzan

AU - Johnson, Alex G.

AU - Rawson, Shaun

AU - Mayer, Megan L.

AU - Doron, Shany

AU - Millman, Adi

AU - Melamed, Sarah

AU - Fedorenko, Taya

AU - Kacen, Assaf

AU - Brandis, Alexander

AU - Mehlman, Tevie

AU - Amitai, Gil

AU - Sorek, Rotem

AU - Kranzusch, Philip J.

PY - 2023/3/2

Y1 - 2023/3/2

N2 - RADAR is a two-protein bacterial defense system that was reported to defend against phage by “editing” messenger RNA. Here, we determine cryo-EM structures of the RADAR defense complex, revealing RdrA as a heptameric, two-layered AAA+ ATPase and RdrB as a dodecameric, hollow complex with twelve surface-exposed deaminase active sites. RdrA and RdrB join to form a giant assembly up to 10 MDa, with RdrA docked as a funnel over the RdrB active site. Surprisingly, our structures reveal an RdrB active site that targets mononucleotides. We show that RdrB catalyzes ATP-to-ITP conversion in vitro and induces the massive accumulation of inosine mononucleotides during phage infection in vivo, limiting phage replication. Our results define ATP mononucleotide deamination as a determinant of RADAR immunity and reveal supramolecular assembly of a nucleotide-modifying machine as a mechanism of anti-phage defense.

AB - RADAR is a two-protein bacterial defense system that was reported to defend against phage by “editing” messenger RNA. Here, we determine cryo-EM structures of the RADAR defense complex, revealing RdrA as a heptameric, two-layered AAA+ ATPase and RdrB as a dodecameric, hollow complex with twelve surface-exposed deaminase active sites. RdrA and RdrB join to form a giant assembly up to 10 MDa, with RdrA docked as a funnel over the RdrB active site. Surprisingly, our structures reveal an RdrB active site that targets mononucleotides. We show that RdrB catalyzes ATP-to-ITP conversion in vitro and induces the massive accumulation of inosine mononucleotides during phage infection in vivo, limiting phage replication. Our results define ATP mononucleotide deamination as a determinant of RADAR immunity and reveal supramolecular assembly of a nucleotide-modifying machine as a mechanism of anti-phage defense.

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U2 - 10.1016/j.cell.2023.01.012

DO - 10.1016/j.cell.2023.01.012

M3 - مقالة

C2 - 36764290

SN - 0092-8674

VL - 186

SP - 987-998.e15

JO - Cell

JF - Cell

IS - 5

ER -

Cryo-EM structure of the RADAR supramolecular anti-phage defense complex

Abstract

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