1. Structure and Activity of the RNA-Targeting Type III-B CRISPR-Cas Complex of Thermus thermophilus
- Author
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Staals, R.H.J., Agari, Y., Maki-Yonekura, S., Zhu, Y., Taylor, D.W., van Duijn, E., Barendregt, A., Vlot, M.C., Koehorst, J.J., Sakamoto, K., Masuda, A., Dohmae, N., Schaap, P.J., Doudna, J.A., Heck, A.J.R., Yonekura, K., van der Oost, J., Shinkai, A., Biomolecular Mass Spectrometry and Proteomics, Sub Biomol.Mass Spectrometry & Proteom., Sub Biomol.Mass Spect. and Proteomics, Biomolecular Mass Spectrometry and Proteomics, Sub Biomol.Mass Spectrometry & Proteom., and Sub Biomol.Mass Spect. and Proteomics
- Subjects
Protein Conformation ,CRISPR-Associated Proteins ,Medical and Health Sciences ,silencing complex ,Protein structure ,Microbiologie ,Models ,Transcription (biology) ,of-flight instrument ,CRISPR ,Systems and Synthetic Biology ,Clustered Regularly Interspaced Short Palindromic Repeats ,Genetics ,Systeem en Synthetische Biologie ,Microscopy ,biology ,Electrospray Ionization ,Bacterial ,High-Throughput Nucleotide Sequencing ,Biological Sciences ,Thermus thermophilus ,antiviral defense ,recognition ,transcription ,Sequence Analysis ,Stereochemistry ,Protein subunit ,interference ,Microbiology ,Electron ,Article ,Structure-Activity Relationship ,Ribonucleases ,Bacterial Proteins ,Molecular Biology ,VLAG ,Trans-activating crRNA ,Spectrometry ,Molecular ,RNA ,Cell Biology ,mass-spectrometry ,Mass ,biology.organism_classification ,Protein Subunits ,immune-system ,escherichia-coli ,CRISPR Loci ,protein ,Developmental Biology - Abstract
The CRISPR-Cas system is a prokaryotic host defense system against genetic elements. The Type III-B CRISPR-Cas system of the bacterium Thermus thermophilus, the TtCmr complex, is composed of six different protein subunits (Cmr1-6) and one crRNA with a stoichiometry of Cmr112131445361:crRNA1. The TtCmr complex copurifies with crRNA species of 40 and 46 nt, originating from a distinct subset of CRISPR loci and spacers. The TtCmr complex cleaves the target RNA at multiple sites with 6 nt intervals via a 5' ruler mechanism. Electron microscopy revealed that the structure of TtCmr resembles a "sea worm" and is composed of a Cmr2-3 heterodimer"tail," a helical backbone of Cmr4 subunits capped by Cmr5 subunits, and a curled "head" containing Cmr1 and Cmr6. Despite having a backbone of only four Cmr4 subunits and being both longer and narrower, the overall architecture of TtCmr resembles that of Type I Cascade complexes.
- Published
- 2013