RNA targeting by the type III-A CRISPR-Cas Csm complex of Thermus thermophilus.

نویسندگان

  • Raymond H J Staals
  • Yifan Zhu
  • David W Taylor
  • Jack E Kornfeld
  • Kundan Sharma
  • Arjan Barendregt
  • Jasper J Koehorst
  • Marnix Vlot
  • Nirajan Neupane
  • Koen Varossieau
  • Keiko Sakamoto
  • Takehiro Suzuki
  • Naoshi Dohmae
  • Shigeyuki Yokoyama
  • Peter J Schaap
  • Henning Urlaub
  • Albert J R Heck
  • Eva Nogales
  • Jennifer A Doudna
  • Akeo Shinkai
  • John van der Oost
چکیده

CRISPR-Cas is a prokaryotic adaptive immune system that provides sequence-specific defense against foreign nucleic acids. Here we report the structure and function of the effector complex of the Type III-A CRISPR-Cas system of Thermus thermophilus: the Csm complex (TtCsm). TtCsm is composed of five different protein subunits (Csm1-Csm5) with an uneven stoichiometry and a single crRNA of variable size (35-53 nt). The TtCsm crRNA content is similar to the Type III-B Cmr complex, indicating that crRNAs are shared among different subtypes. A negative stain EM structure of the TtCsm complex exhibits the characteristic architecture of Type I and Type III CRISPR-associated ribonucleoprotein complexes. crRNA-protein crosslinking studies show extensive contacts between the Csm3 backbone and the bound crRNA. We show that, like TtCmr, TtCsm cleaves complementary target RNAs at multiple sites. Unlike Type I complexes, interference by TtCsm does not proceed via initial base pairing by a seed sequence.

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عنوان ژورنال:
  • Molecular cell

دوره 56 4  شماره 

صفحات  -

تاریخ انتشار 2014