The CRISPR ancillary effector Can2 is a dual-specificity nuclease potentiating type III CRISPR defence

Wenlong Zhu; Stuart John McQuarrie; Sabine Gruschow; Stephen McMahon; Shirley Graham; Tracey Gloster; Malcolm White

doi:10.1093/nar/gkab073

The CRISPR ancillary effector Can2 is a dual-specificity nuclease potentiating type III CRISPR defence

Wenlong Zhu, Stuart John McQuarrie, Sabine Gruschow, Stephen McMahon, Shirley Graham, Tracey Gloster^*, Malcolm White^*

^*Corresponding author for this work

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

Abstract

Cells and organisms have a wide range of mechanisms to defend against infection by viruses and other mobile genetic elements (MGE). Type III CRISPR systems detect foreign RNA and typically generate cyclic oligoadenylate (cOA) second messengers that bind to ancillary proteins with CARF (CRISPR associated Rossman fold) domains. This results in the activation of fused effector domains for antiviral defence. The best characterised CARF family effectors are the Csm6/Csx1 ribonucleases and DNA nickase Can1. Here we investigate a widely distributed CARF family effector with a nuclease domain, which we name Can2 (CRISPR ancillary nuclease 2). Can2 is activated by cyclic tetra-adenylate (cA₄) and displays both DNase and RNase activity, providing effective immunity against plasmid transformation and bacteriophage infection in Escherichia coli. The structure of Can2 in complex with cA₄ suggests a mechanism for the cA₄-mediated activation of the enzyme, whereby an active site cleft is exposed on binding the activator. These findings extend our understanding of type III CRISPR cOA signalling and effector function.

Original language	English
Article number	gkab073
Pages (from-to)	2777–2789
Number of pages	13
Journal	Nucleic Acids Research
Volume	49
Issue number	5
Early online date	15 Feb 2021
DOIs	https://doi.org/10.1093/nar/gkab073
Publication status	Published - 18 Mar 2021

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10.1093/nar/gkab073Licence: CC BY

Zhu_2021_NAR_CRISPR-ancillary_CC
Copyright The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
Final published version, 2.89 MBLicence: CC BY

Dissecting Molecular Biology of Cyclic: Dissecting the Molecular Biology of Cyclic Oligoadenylate Signalling
White, M. (PI) & Gloster, T. (CoI)
BBSRC
1/01/20 → 31/12/23
Project: Standard
ISSF3 Round 4 _ Stuart McQuarrie: ISSF3 funding round 4
Gloster, T. (PI) & White, M. (CoI)
1/02/19 → 31/08/19
Project: Standard
Cyclic oligoadenylate signalling: Cyclic oligoadenylate signalling - a new type of antiviral response
White, M. (PI)
1/01/19 → 31/12/21
Project: Standard

Structure of Can2 from Sulfobacillus thermosulfidooxidans in complex with cyclic tetra-adenylate (cA4) (dataset)
McQuarrie, S. J. (Creator), McMahon, S. (Creator), Gloster, T. (Creator), White, M. (Creator), Graham, S. (Creator), Zhu, W. (Creator) & Gruschow, S. (Creator), Protein Data Bank (PDB), 2021
DOI: 10.2210/pdb7bdv/pdb, https://www.rcsb.org/structure/7BDV
Dataset

Cite this

@article{2865bdf7be594b46a15558951db4c3fb,

title = "The CRISPR ancillary effector Can2 is a dual-specificity nuclease potentiating type III CRISPR defence",

abstract = "Cells and organisms have a wide range of mechanisms to defend against infection by viruses and other mobile genetic elements (MGE). Type III CRISPR systems detect foreign RNA and typically generate cyclic oligoadenylate (cOA) second messengers that bind to ancillary proteins with CARF (CRISPR associated Rossman fold) domains. This results in the activation of fused effector domains for antiviral defence. The best characterised CARF family effectors are the Csm6/Csx1 ribonucleases and DNA nickase Can1. Here we investigate a widely distributed CARF family effector with a nuclease domain, which we name Can2 (CRISPR ancillary nuclease 2). Can2 is activated by cyclic tetra-adenylate (cA4) and displays both DNase and RNase activity, providing effective immunity against plasmid transformation and bacteriophage infection in Escherichia coli. The structure of Can2 in complex with cA4 suggests a mechanism for the cA4-mediated activation of the enzyme, whereby an active site cleft is exposed on binding the activator. These findings extend our understanding of type III CRISPR cOA signalling and effector function.",

author = "Wenlong Zhu and McQuarrie, {Stuart John} and Sabine Gruschow and Stephen McMahon and Shirley Graham and Tracey Gloster and Malcolm White",

note = "Funding: Biotechnology and Biological Sciences Research Council [BB/S000313/1 to M.F.W., BB/T004789/1 to M.F.W. and T.M.G.]; Wellcome Trust Institutional Strategic Support Funding [204821/Z/16/Z to M.F.W. and T.M.G.]; China Scholarship Council [201703780015 to W.Z.]. Funding for open access charge: RCUK block grant.",

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doi = "10.1093/nar/gkab073",

language = "English",

volume = "49",

pages = "2777–2789",

journal = "Nucleic Acids Research",

issn = "0305-1048",

publisher = "Oxford University Press",

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AU - Zhu, Wenlong

AU - McQuarrie, Stuart John

AU - Gruschow, Sabine

AU - McMahon, Stephen

AU - Graham, Shirley

AU - Gloster, Tracey

AU - White, Malcolm

N1 - Funding: Biotechnology and Biological Sciences Research Council [BB/S000313/1 to M.F.W., BB/T004789/1 to M.F.W. and T.M.G.]; Wellcome Trust Institutional Strategic Support Funding [204821/Z/16/Z to M.F.W. and T.M.G.]; China Scholarship Council [201703780015 to W.Z.]. Funding for open access charge: RCUK block grant.

PY - 2021/3/18

Y1 - 2021/3/18

N2 - Cells and organisms have a wide range of mechanisms to defend against infection by viruses and other mobile genetic elements (MGE). Type III CRISPR systems detect foreign RNA and typically generate cyclic oligoadenylate (cOA) second messengers that bind to ancillary proteins with CARF (CRISPR associated Rossman fold) domains. This results in the activation of fused effector domains for antiviral defence. The best characterised CARF family effectors are the Csm6/Csx1 ribonucleases and DNA nickase Can1. Here we investigate a widely distributed CARF family effector with a nuclease domain, which we name Can2 (CRISPR ancillary nuclease 2). Can2 is activated by cyclic tetra-adenylate (cA4) and displays both DNase and RNase activity, providing effective immunity against plasmid transformation and bacteriophage infection in Escherichia coli. The structure of Can2 in complex with cA4 suggests a mechanism for the cA4-mediated activation of the enzyme, whereby an active site cleft is exposed on binding the activator. These findings extend our understanding of type III CRISPR cOA signalling and effector function.

AB - Cells and organisms have a wide range of mechanisms to defend against infection by viruses and other mobile genetic elements (MGE). Type III CRISPR systems detect foreign RNA and typically generate cyclic oligoadenylate (cOA) second messengers that bind to ancillary proteins with CARF (CRISPR associated Rossman fold) domains. This results in the activation of fused effector domains for antiviral defence. The best characterised CARF family effectors are the Csm6/Csx1 ribonucleases and DNA nickase Can1. Here we investigate a widely distributed CARF family effector with a nuclease domain, which we name Can2 (CRISPR ancillary nuclease 2). Can2 is activated by cyclic tetra-adenylate (cA4) and displays both DNase and RNase activity, providing effective immunity against plasmid transformation and bacteriophage infection in Escherichia coli. The structure of Can2 in complex with cA4 suggests a mechanism for the cA4-mediated activation of the enzyme, whereby an active site cleft is exposed on binding the activator. These findings extend our understanding of type III CRISPR cOA signalling and effector function.

U2 - 10.1093/nar/gkab073

DO - 10.1093/nar/gkab073

M3 - Article

C2 - 33590098

AN - SCOPUS:85103229027

SN - 0305-1048

VL - 49

SP - 2777

EP - 2789

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 5

M1 - gkab073

ER -

The CRISPR ancillary effector Can2 is a dual-specificity nuclease potentiating type III CRISPR defence

Abstract

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Projects

Dissecting Molecular Biology of Cyclic: Dissecting the Molecular Biology of Cyclic Oligoadenylate Signalling

ISSF3 Round 4 _ Stuart McQuarrie: ISSF3 funding round 4

Cyclic oligoadenylate signalling: Cyclic oligoadenylate signalling - a new type of antiviral response

Datasets

Structure of Can2 from Sulfobacillus thermosulfidooxidans in complex with cyclic tetra-adenylate (cA4) (dataset)

Cite this