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

Staals, R. H.; Zhu, Y.; Taylor, D. W.; Kornfeld, J. E.; Sharma, K.; Barendregt, A.; Koehorst, J. F.; Vlot, M.; Neupane, N.; Varossieau, K.; Sakamoto, K.; Suzuki, T.; Dohmae, N.; Yokoyama, S.; Schaap, P. J.; Urlaub, H.; Heck, A. J. R.; Nogales, E.; Doudna, J. A.; Shinkai, A.; van der Oost, J.

doi:10.1016/j.molcel.2014.10.005

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学術論文

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

MPS-Authors

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Sharma, K.
Research Group of Bioanalytical Mass Spectrometry, MPI for Biophysical Chemistry, Max Planck Society;

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Urlaub, H.
Research Group of Bioanalytical Mass Spectrometry, MPI for Biophysical Chemistry, Max Planck Society;

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http://www.sciencedirect.com/science/article/pii/S1097276514007898/pdfft?md5=50f71ae832e0c62971b07b5ecfeb9b5c&pid=1-s2.0-S1097276514007898-main.pdf
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引用

Staals, R. H., Zhu, Y., Taylor, D. W., Kornfeld, J. E., Sharma, K., Barendregt, A., Koehorst, J. F., Vlot, M., Neupane, N., Varossieau, K., Sakamoto, K., Suzuki, T., Dohmae, N., Yokoyama, S., Schaap, P. J., Urlaub, H., Heck, A. J. R., Nogales, E., Doudna, J. A., Shinkai, A., & van der Oost, J. (2014). RNA targeting by the type III-A CRISPR-Cas Csm complex of Thermus thermophilus. Molecular Cell, 56(4), 518-530. doi:10.1016/j.molcel.2014.10.005.

引用: https://hdl.handle.net/11858/00-001M-0000-0024-63C9-B

要旨

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.