1. Structure and Activity of the RNA-Targeting Type III-B CRISPR-Cas Complex of Thermus thermophilus
- Author
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Staals, Raymond HJ, Agari, Yoshihiro, Maki-Yonekura, Saori, Zhu, Yifan, Taylor, David W, van Duijn, Esther, Barendregt, Arjan, Vlot, Marnix, Koehorst, Jasper J, Sakamoto, Keiko, Masuda, Akiko, Dohmae, Naoshi, Schaap, Peter J, Doudna, Jennifer A, Heck, Albert JR, Yonekura, Koji, van der Oost, John, and Shinkai, Akeo
- Subjects
Biochemistry and Cell Biology ,Biological Sciences ,Bacterial Proteins ,CRISPR-Associated Proteins ,Clustered Regularly Interspaced Short Palindromic Repeats ,High-Throughput Nucleotide Sequencing ,Microscopy ,Electron ,Models ,Molecular ,Protein Conformation ,Protein Subunits ,RNA ,Bacterial ,Ribonucleases ,Sequence Analysis ,RNA ,Spectrometry ,Mass ,Electrospray Ionization ,Structure-Activity Relationship ,Thermus thermophilus ,Medical and Health Sciences ,Developmental Biology ,Biological sciences ,Biomedical and clinical sciences ,Health sciences - 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