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Cited by 25CRISPR and Genetic EngineeringBacterial Genetics and BiotechnologyBacteriophages and microbial interactions

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CRISPR antiphage defence mediated by the cyclic nucleotide-binding membrane protein Csx23

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Grüschow, Sabine, McQuarrie, Stuart, Ackermann, Katrin, McMahon, Stephen, Bode, Bela E, Gloster, Tracey M, White, Malcolm F·2024·Nucleic Acids Research
DOI 10.1093/nar/gkae167PMC PMC11014256

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Abstract

CRISPR-Cas provides adaptive immunity in prokaryotes. Type III CRISPR systems detect invading RNA and activate the catalytic Cas10 subunit, which generates a range of nucleotide second messengers to signal infection. These molecules bind and activate a diverse range of effector proteins that provide immunity by degrading viral components and/or by disturbing key aspects of cellular metabolism to slow down viral replication. Here, we focus on the uncharacterised effector Csx23, which is widespread in Vibrio cholerae. Csx23 provides immunity against plasmids and phage when expressed in Escherichia coli along with its cognate type III CRISPR system. The Csx23 protein localises in the membrane using an N-terminal transmembrane α-helical domain and has a cytoplasmic C-terminal domain that binds cyclic tetra-adenylate (cA4), activating its defence function. Structural studies reveal a tetrameric structure with a novel fold that binds cA4 specifically. Using pulse EPR, we demonstrate that cA4 binding to the cytoplasmic domain of Csx23 results in a major perturbation of the transmembrane domain, consistent with the opening of a pore and/or disruption of membrane integrity. This work reveals a new class of cyclic nucleotide binding protein and provides key mechanistic detail on a membrane-associated CRISPR effector.

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Metadata

Authors
Grüschow, S., McQuarrie, S., Ackermann, K., McMahon, S., Bode, B. E., Gloster, T. M., & White, M. F.
Affiliations
University of St Andrews (GB)
Corresponding author
  • Gloster, Tracey M
  • White, Malcolm F
ORCID
Venue
Nucleic Acids Research
Publisher
Oxford University Press (OUP)
Published
2024-03-13
Volume
52
Issue
6
Pages
2761-2775
ISSN
  • 0305-1048 (ISSN-L)
  • 1362-4962
DOI
10.1093/nar/gkae167
PMC
PMC11014256
OpenAlex
W4392722047
Cited by
25
References
1
Citations by year
2026: 10 · 2025: 11 · 2024: 4
FWCI
3.40
Type
journal article
Language
en
Open access
Licence
Locations
  • Nucleic Acids Research (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • St Andrews Research Repository (St Andrews Research Repository) (submittedVersion, cc-by)
Funders
  • Biotechnology and Biological Sciences Research Council (BB/T004789/1)
  • European Research Council Advanced (101018608)
  • Engineering and Physical Sciences Research Council (EP/X016455/1)
  • BBSRC (BB/R013780/1)
  • University of St Andrews
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Immunology and Microbiology › Virology › Bacteriophages and microbial interactions
Keywords
membrane proteins, Vibrio cholerae, transmembrane domain, protein structure, pulsed EPR, prokaryotic adaptive immunity
MeSH terms
Adenine Nucleotides, Bacterial Proteins, Membrane Proteins, Nucleotides, Cyclic, Vibrio cholerae, Second Messenger Systems, CRISPR-Cas Systems, CRISPR-Associated Proteins

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