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Cited by 17CRISPR and Genetic EngineeringBacterial Genetics and BiotechnologyBacteriophages and microbial interactions

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Filament formation activates protease and ring nuclease activities of CRISPR Lon-SAVED

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Smalakyte, Dalia, Ruksenaite, Audrone, Sasnauskas, Giedrius, Tamulaitiene, Giedre, Tamulaitis, Gintautas·2024·Molecular Cell
DOI 10.1016/j.molcel.2024.09.002

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Abstract

To combat phage infection, type III CRISPR-Cas systems utilize cyclic oligoadenylates (cA n) signaling to activate various auxiliary effectors, including the CRISPR-associated Lon-SAVED protease CalpL, which forms a tripartite effector system together with an anti-σ factor, CalpT, and an ECF-like σ factor, CalpS. Here, we report the characterization of the Candidatus Cloacimonas acidaminovorans CalpL-CalpT-CalpS. We demonstrate that cA 4 binding triggers CalpL filament formation and activates it to cleave CalpT within the CalpT-CalpS dimer. This cleavage exposes the CalpT C-degron, which targets it for further degradation by cellular proteases. Consequently, CalpS is released to bind to RNA polymerase, causing growth arrest in E. coli. Furthermore, the CalpL-CalpT-CalpS system is regulated by the SAVED domain of CalpL, which is a ring nuclease that cleaves cA 4 in a sequential three-step mechanism. These findings provide key mechanistic details for the activation, proteolytic events, and regulation of the signaling cascade in the type III CRISPR-Cas immunity.

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Metadata

Authors
Smalakyte, D., Ruksenaite, A., Sasnauskas, G., Tamulaitiene, G., & Tamulaitis, G.
Affiliations
Vilnius University (LT)
Corresponding author
Tamulaitis, Gintautas
ORCID
Venue
Molecular Cell
Publisher
Elsevier BV
Published
2024-11
Volume
84
Issue
21
Pages
4239-4255.e8
ISSN
1097-2765 (ISSN-L)
DOI
10.1016/j.molcel.2024.09.002
OpenAlex
W4403065814
Cited by
17
References
5
Citations by year
2026: 6 · 2025: 10 · 2024: 1
FWCI
2.32
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Cell (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
Lietuvos Mokslo Taryba (S-MIP-22-09)
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
type III CRISPR-Cas, CALP, cyclic oligoadenylate, filamentation, anti-sigma factor, ECF sigma factor, phage immunity, RNA polymerase binding, growth arrest
MeSH terms
Bacterial Proteins, Escherichia coli, Protein Binding, Sigma Factor, Signal Transduction, Gene Expression Regulation, Bacterial, Proteolysis, CRISPR-Cas Systems, CRISPR-Associated Proteins

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