|Entry||Database: EMDB / ID: 3454|
|Title||RNA activation-independent DNA targeting by a Csm complex of the Type III CRISPR-Cas system|
|Map data||Csm complex with crRNA from T.onnurineus|
|Sample||Csm complex with crRNA from T.onnurineus|
|Source||Thermococcus onnurineus na1 / / archaea / thermophilic|
|Method||single particle reconstruction / 25 Å resolution|
|Authors||Jung TY / Song JJ / Hebert H / Woo EJ / Oh BH / Park KH / An Y|
|Citation||Journal: EMBO Rep. / Year: 2017|
Title: RNA activation-independent DNA targeting of the Type III CRISPR-Cas system by a Csm complex.
Authors: Kwang-Hyun Park / Yan An / Tae-Yang Jung / In-Young Baek / Haemin Noh / Woo-Chan Ahn / Hans Hebert / Ji-Joon Song / Jeong-Hoon Kim / Byung-Ha Oh / Eui-Jeon Woo
Abstract: The CRISPR-Cas system is an adaptive and heritable immune response that destroys invading foreign nucleic acids. The effector complex of the Type III CRISPR-Cas system targets RNA and DNA in a ...The CRISPR-Cas system is an adaptive and heritable immune response that destroys invading foreign nucleic acids. The effector complex of the Type III CRISPR-Cas system targets RNA and DNA in a transcription-coupled manner, but the exact mechanism of DNA targeting by this complex remains elusive. In this study, an effector Csm holocomplex derived from is reconstituted with a minimalistic combination of Csm12345, and shows RNA targeting and RNA-activated single-stranded DNA (ssDNA) targeting activities. Unexpectedly, in the absence of an RNA transcript, it cleaves ssDNA containing a sequence complementary to the bound crRNA guide region in a manner dependent on the HD domain of the Csm1 subunit. This nuclease activity is blocked by a repeat tag found in the host CRISPR loci. The specific cleavage of ssDNA without a target RNA suggests a novel ssDNA targeting mechanism of the Type III system, which could facilitate the efficient and complete degradation of foreign nucleic acids.
Copyright: 2017 The Authors.
|Date||Deposition: Nov 2, 2016 / Header (metadata) release: Dec 7, 2016 / Map release: Apr 5, 2017 / Last update: Aug 2, 2017|
Downloads & links
|File||emd_3454.map.gz (map file in CCP4 format, 19653 KB)|
|Projections & slices|
Images are generated by Spider package.
|Voxel size||X=Y=Z: 2.07 Å|
CCP4 map header:
-Entire Csm complex with crRNA from T.onnurineus
|Entire||Name: Csm complex with crRNA from T.onnurineus / Number of components: 1|
|Mass||Experimental: 280 kDa|
-Component #1: protein, Csm complex with crRNA from T.onnurineus
|Specimen||Specimen state: particle|
|Sample solution||Specimen conc.: 0.025 mg/ml / pH: 8|
|Vitrification||Cryogen name: NONE|
-Electron microscopy imaging
|Imaging||Microscope: JEOL 2100F|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Electron dose: 30 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Cs: 2 mm / Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: JEOL|
|Camera||Detector: TVIPS TEMCAM-F415 (4k x 4k)|
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