RRC ID 50343
Author Staals RHJ, Agari Y, Maki-Yonekura S, Zhu Y, Taylor DW, van Duijn E, Barendregt A, Vlot M, Koehorst JJ, Sakamoto K, Masuda A, Dohmae N, Schaap PJ, Doudna JA, Heck AJR, Yonekura K, van der Oost J, Shinkai A.
Title Structure and activity of the RNA-targeting Type III-B CRISPR-Cas complex of Thermus thermophilus.
Journal Mol. Cell
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.
Volume 52(1)
Pages 135-145
Published 2013-10-10
DOI 10.1016/j.molcel.2013.09.013
PMID 24119403
PMC PMC4006948
MeSH Bacterial Proteins / chemistry Bacterial Proteins / genetics Bacterial Proteins / metabolism* CRISPR-Associated Proteins / chemistry CRISPR-Associated Proteins / genetics CRISPR-Associated Proteins / metabolism* Clustered Regularly Interspaced Short Palindromic Repeats High-Throughput Nucleotide Sequencing Microscopy, Electron Models, Molecular Protein Conformation Protein Subunits RNA, Bacterial / chemistry RNA, Bacterial / genetics RNA, Bacterial / metabolism* Ribonucleases / chemistry Ribonucleases / genetics Ribonucleases / metabolism* Sequence Analysis, RNA Spectrometry, Mass, Electrospray Ionization Structure-Activity Relationship Thermus thermophilus / genetics Thermus thermophilus / metabolism*
IF 14.248
Resource
DNA material pUC-cmr6h (RDB15775) PC014162-42 (RDB15779).