ArticleThe Journal of biological chemistry2014
A complex of Cas proteins 5, 6, and 7 is required for the biogenesis and stability of clustered regularly interspaced short palindromic repeats (crispr)-derived rnas (crrnas) in Haloferax volcanii.
Article in The Journal of biological chemistry, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 55 papers, 1 of them a synthesis that pooled it.
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Who cites it
55 citing papers in PubMed, 1 synthesis or guideline pooled it, 83 citations in OpenAlex.
- CRISPR/Cas-Based Modifications for Therapeutic Applications: A Review.Molecular biotechnology · 2022Pooled it
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- Archaeal RNA processing and regulation: expanding the functional landscape.Microbiology and molecular biology reviews : MMBR · 2025Review
- The stress of carrying CRISPR-Cas.Virulence · 2025Article
- Decoding the Chemical Language of Ribosomally Synthesized and Post-Translationally Modified Peptides from the Untapped Archaea Domain.Angewandte Chemie (International ed. in English) · 2025Article
- CRISPR-Cas induced self-targeting identifies key players in archaeal microhomology-mediated end joining.microLife · 2025Review
- Internal in-frame translation generates Cas11b, which is important for effective interference in an archaeal CRISPR-Cas system.Frontiers in microbiology · 2025Article
- Biochemical plasticity of the Escherichia coli CRISPR Cascade revealed by in vitro reconstitution of Cascade activities from purified Cas proteins.The FEBS journal · 2024Article
- Article
- Article
- Cold Cas: reevaluating the occurrence of CRISPR/Cas systems inFrontiers in microbiology · 2023Article
- CRISPR Interference as a Tool to Repress Gene Expression in Haloferax volcanii.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Insights into synthesis and function of KsgA/Dim1-dependent rRNA modifications in archaea.Nucleic acids research · 2021Article
- Expansion of the CRISPR/Cas Genome-Sculpting Toolbox: Innovations, Applications and Challenges.Molecular diagnosis & therapy · 2021Review
- A Small RNA Is Linking CRISPR-Cas and Zinc Transport.Frontiers in molecular biosciences · 2021Article
- Genome editing technologies: CRISPR, LEAPER, RESTORE, ARCUT, SATI, and RESCUE.EXCLI journal · 2021Review
- Heavily Armed Ancestors: CRISPR Immunity and Applications in Archaea with a Comparative Analysis of CRISPR Types in Sulfolobales.Biomolecules · 2020Review
- Adaptation induced by self-targeting in a type I-B CRISPR-Cas system.The Journal of biological chemistry · 2020Article
- The switch complex ArlCDE connects the chemotaxis system and the archaellum.Molecular microbiology · 2020Article
- Interaction of two strongly divergent archaellins stabilizes the structure of the Halorubrum archaellum.MicrobiologyOpen · 2020Article
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Authors and funding
15 authors at 7 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The clustered regularly interspaced short palindromic repeats/CRISPR-associated (CRISPR-Cas) system is a prokaryotic defense mechanism against foreign genetic elements. A plethora of CRISPR-Cas versions exist, with more than 40 different Cas protein families and several different molecular approaches to fight the invading DNA. One of the key players in the system is the CRISPR-derived RNA (crRNA), which directs the invader-degrading Cas protein complex to the invader. The CRISPR-Cas types I and III use the Cas6 protein to generate mature crRNAs. Here, we show that the Cas6 protein is necessary for crRNA production but that additional Cas proteins that form a CRISPR-associated complex for antiviral defense (Cascade)-like complex are needed for crRNA stability in the CRISPR-Cas type I-B system in Haloferax volcanii in vivo. Deletion of the cas6 gene results in the loss of mature crRNAs and interference. However, cells that have the complete cas gene cluster (cas1-8b) removed and are transformed with the cas6 gene are not able to produce and stably maintain mature crRNAs. crRNA production and stability is rescued only if cas5, -6, and -7 are present. Mutational analysis of the cas6 gene reveals three amino acids (His-41, Gly-256, and Gly-258) that are essential for pre-crRNA cleavage, whereas the mutation of two amino acids (Ser-115 and Ser-224) leads to an increase of crRNA amounts. This is the first systematic in vivo analysis of Cas6 protein variants. In addition, we show that the H. volcanii I-B system contains a Cascade-like complex with a Cas7, Cas5, and Cas6 core that protects the crRNA.
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