RNA Targeting by the Type III-A CRISPR-Cas Csm Complex of Thermus thermophilus

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

doi:10.1016/j.molcel.2014.10.005

RNA Targeting by the Type III-A CRISPR-Cas Csm Complex of Thermus thermophilus

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 ShinkaiJohn van der Oost

Research output: Contribution to journal › Article › Academic › peer-review

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.

Original language	English
Pages (from-to)	518-30
Number of pages	13
Journal	Molecular Cell
Volume	56
Issue number	4
DOIs	https://doi.org/10.1016/j.molcel.2014.10.005
Publication status	Published - 20 Nov 2014

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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., Dohmae, N., Yokoyama, S., Schaap, P. J., Urlaub, H., Heck, A. J. R., Nogales, E., Doudna, J. A., ... van der Oost, J. (2014). RNA Targeting by the Type III-A CRISPR-Cas Csm Complex of Thermus thermophilus. Molecular Cell, 56(4), 518-30. https://doi.org/10.1016/j.molcel.2014.10.005

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title = "RNA Targeting by the Type III-A CRISPR-Cas Csm Complex of Thermus thermophilus",

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.",

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

year = "2014",

month = nov,

day = "20",

doi = "10.1016/j.molcel.2014.10.005",

language = "English",

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Staals, RHJ, Zhu, Y, Taylor, DW, Kornfeld, JE, Sharma, K, Barendregt, A, Koehorst, JJ, Vlot, M, Neupane, N, Varossieau, K, Sakamoto, K, Suzuki, T, Dohmae, N, Yokoyama, S, Schaap, PJ, Urlaub, H, Heck, AJR, Nogales, E, Doudna, JA, Shinkai, A & van der Oost, J 2014, 'RNA Targeting by the Type III-A CRISPR-Cas Csm Complex of Thermus thermophilus', Molecular Cell, vol. 56, no. 4, pp. 518-30. https://doi.org/10.1016/j.molcel.2014.10.005

TY - JOUR

T1 - RNA Targeting by the Type III-A CRISPR-Cas Csm Complex of Thermus thermophilus

AU - Staals, Raymond H J

AU - Zhu, Yifan

AU - Taylor, David W

AU - Kornfeld, Jack E

AU - Sharma, Kundan

AU - Barendregt, Arjan

AU - Koehorst, Jasper J

AU - Vlot, Marnix

AU - Neupane, Nirajan

AU - Varossieau, Koen

AU - Sakamoto, Keiko

AU - Suzuki, Takehiro

AU - Dohmae, Naoshi

AU - Yokoyama, Shigeyuki

AU - Schaap, Peter J

AU - Urlaub, Henning

AU - Heck, Albert J R

AU - Nogales, Eva

AU - Doudna, Jennifer A

AU - Shinkai, Akeo

AU - van der Oost, John

PY - 2014/11/20

Y1 - 2014/11/20

N2 - 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.

AB - 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.

U2 - 10.1016/j.molcel.2014.10.005

DO - 10.1016/j.molcel.2014.10.005

M3 - Article

C2 - 25457165

SN - 1097-2765

VL - 56

SP - 518

EP - 530

JO - Molecular Cell

JF - Molecular Cell

IS - 4

ER -

RNA Targeting by the Type III-A CRISPR-Cas Csm Complex of Thermus thermophilus

Abstract

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