Structure and Activity of the RNA-Targeting Type III-B CRISPR-Cas Complex of Thermus thermophilus

Raymond H.J. Staals, Yoshihiro Agari, Saori Maki-Yonekura, Yifan Zhu, David W. Taylor, Esther vanDuijn, Arjan Barendregt, Marnix Vlot, Jasper J. Koehorst, Keiko Sakamoto, Akiko Masuda, Naoshi Dohmae, Peter J. Schaap, Jennifer A. Doudna, Albert J.R. Heck, Koji Yonekura, John Van der Oost, Akeo Shinkai

Research output: Contribution to journalArticlepeer-review

149 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)135-145
Number of pages11
JournalMolecular Cell
Volume52
Issue number1
DOIs
Publication statusPublished - 2013 Oct 10
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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