ArticleRNA biology2019
The ribonuclease activity of Csm6 is required for anti-plasmid immunity by Type III-A CRISPR-Cas systems.
Article in RNA biology, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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Who cites it
43 citing papers in PubMed, 73 citations in OpenAlex.
- Cas10 residues lining the target RNA binding channel regulate interference by distinguishing cognate target RNA from mismatched targets.RNA biology · 2026Article
- A phage-encoded anti-CRISPR protein co-opts host enolase to prevent type III CRISPR immunity.Nature microbiology · 2025Article
- The extended mobility of plasmids.Nucleic acids research · 2025Review
- CARF-dependent preferential RNA cleavage by Csm6 increases drug susceptibility of mycobacteria.Nucleic acids research · 2025Article
- Selective degradation of phage RNAs by the Csm6 ribonuclease provides robust type III CRISPR immunity in Streptococcus thermophilus.Nucleic acids research · 2024Article
- Molecular basis for cA6 synthesis by a type III-A CRISPR-Cas enzyme and its conversion to cA4 production.Nucleic acids research · 2024Article
- Retention of the RNA ends provides the molecular memory for maintaining the activation of the Csm complex.Nucleic acids research · 2024Article
- Substrate selectivity and catalytic activation of the type III CRISPR ancillary nuclease Can2.Nucleic acids research · 2024Article
- Current Updates of CRISPR/Cas System and Anti-CRISPR Proteins: Innovative Applications to Improve the Genome Editing Strategies.International journal of nanomedicine · 2024Review
- Article
- The influence of the copy number of invader on the fate of bacterial host cells in the antiviral defense by CRISPR-Cas10 DNases.Engineering microbiology · 2023Article
- Activation of Csm6 ribonuclease by cyclic nucleotide binding: in an emergency, twist to open.Nucleic acids research · 2023Article
- An Endogenous Staphylococcus aureus CRISPR-Cas System Limits Phage Proliferation and Is Efficiently Excised from the Genome as Part of the SCCMicrobiology spectrum · 2023Article
- EspB and HtpG interact with the type III-A CRISPR/Cas system ofFrontiers in molecular biosciences · 2023Article
- The diverse arsenal of type III CRISPR-Cas-associated CARF and SAVED effectors.Biochemical Society transactions · 2022Review
- Type III-A CRISPR systems as a versatile gene knockdown technology.RNA (New York, N.Y.) · 2022Article
- Inactivation of Target RNA Cleavage of a III-B CRISPR-Cas System Induces Robust Autoimmunity inInternational journal of molecular sciences · 2022Article
- Structural and biochemical characterization of in vivo assembled Lactococcus lactis CRISPR-Csm complex.Communications biology · 2022Article
- Unique properties of spacer acquisition by the type III-A CRISPR-Cas system.Nucleic acids research · 2022Article
- Enzymatic properties of CARF-domain proteins inFrontiers in microbiology · 2022Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
CRISPR-Cas systems provide prokaryotes with RNA-based adaptive immunity against viruses and plasmids. A unique feature of Type III CRISPR-Cas systems is that they selectively target transcriptionally-active invader DNA, and can cleave both the expressed RNA transcripts and source DNA. The Type III-A effector crRNP (CRISPR RNA-Cas protein complex), which contains Cas proteins Csm1-5, recognizes and degrades invader RNA and DNA in a crRNA-guided, manner. Interestingly, Type III-A systems also employ Csm6, an HEPN family ribonuclease that does not stably associate with the Type III-A effector crRNP, but nevertheless contributes to defense via mechanistic details that are still being determined. Here, we investigated the mechanism of action of Csm6 in Type III-A CRISPR-Cas systems from Lactococcus lactis, Staphylococcus epidermidis, and Streptococcus thermophilus expressed in Escherichia coli. We found that L. lactis and S. epidermidis Csm6 cleave RNA specifically after purines in vitro, similar to the activity reported for S. thermophilus Csm6. Moreover, L. lactis Csm6 functions as a divalent metal-independent, single strand-specific endoribonuclease that depends on the conserved HEPN domain. In vivo, we show that deletion of csm6 or expression of an RNase-defective form of Csm6 disrupts crRNA-dependent loss of plasmid DNA in all three systems expressed in E. coli. Mutations in the Csm1 palm domain, which are known to deactivate Csm6 ribonuclease activity, also prevent plasmid loss in the three systems. The results indicate that Csm6 ribonuclease activity rather than Csm1-mediated DNase activity effects anti-plasmid immunity by the three Type III-A systems investigated.
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