Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

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Record number 480592
Title RNA Targeting by the Type III-A CRISPR-Cas Csm Complex of Thermus thermophilus
Author(s) Staals, R.H.J.; Zhu, Y.; Taylor, D.W.; Kornfeld, J.E.; Sharma, K.; Barendregt, A.; Koehorst, J.J.; Vlot, M.; Neupane, N.; Varossieau, K.; Sakamoto, K.; Suzuki, T.; Schaap, P.J.; Urlaub, H.; Heck, A.J.R.; Nogales, E.; Doudna, J.A.; Shinkai, A.; Oost, J. van der
Source Molecular Cell 56 (2014)4. - ISSN 1097-2765 - p. 518 - 530.
DOI http://dx.doi.org/10.1016/j.molcel.2014.10.005
Department(s) Microbiological Laboratory
Systems and Synthetic Biology
Laboratory of Nematology
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2014
Keyword(s) guided surveillance complex - bacterial immune-system - adaptive immunity - mass-spectrometry - crystal-structure - escherichia-coli - haloferax-volcanii - antiviral defense - seed sequence - protein
Abstract 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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