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Yorodumi- EMDB-29039: Cryo-EM structure of Cascade-DNA-TniQ-TnsC complex (composite) in... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-29039 | |||||||||
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Title | Cryo-EM structure of Cascade-DNA-TniQ-TnsC complex (composite) in type I-B CAST system | |||||||||
Map data | ||||||||||
Sample |
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Keywords | CRISPR / transposon / TnsC / TniQ / DNA BINDING PROTEIN | |||||||||
Function / homology | Function and homology information | |||||||||
Biological species | Peltigera membranacea (fungus) / Nostoc sp. 'Peltigera membranacea cyanobiont' 210A (bacteria) / synthetic construct (others) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||
Authors | Chang L / Wang S | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Cell / Year: 2023 Title: Molecular mechanism for Tn7-like transposon recruitment by a type I-B CRISPR effector. Authors: Shukun Wang / Clinton Gabel / Romana Siddique / Thomas Klose / Leifu Chang / Abstract: Tn7-like transposons have co-opted CRISPR-Cas systems to facilitate the movement of their own DNA. These CRISPR-associated transposons (CASTs) are promising tools for programmable gene knockin. A key ...Tn7-like transposons have co-opted CRISPR-Cas systems to facilitate the movement of their own DNA. These CRISPR-associated transposons (CASTs) are promising tools for programmable gene knockin. A key feature of CASTs is their ability to recruit Tn7-like transposons to nuclease-deficient CRISPR effectors. However, how Tn7-like transposons are recruited by diverse CRISPR effectors remains poorly understood. Here, we present the cryo-EM structure of a recruitment complex comprising the Cascade complex, TniQ, TnsC, and the target DNA in the type I-B CAST from Peltigera membranacea cyanobiont 210A. Target DNA recognition by Cascade induces conformational changes in Cas6 and primes TniQ recruitment through its C-terminal domain. The N-terminal domain of TniQ is bound to the seam region of the TnsC spiral heptamer. Our findings provide insights into the diverse mechanisms for the recruitment of Tn7-like transposons to CRISPR effectors and will aid in the development of CASTs as gene knockin tools. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_29039.map.gz | 16 MB | EMDB map data format | |
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Header (meta data) | emd-29039-v30.xml emd-29039.xml | 23.4 KB 23.4 KB | Display Display | EMDB header |
Images | emd_29039.png | 116 KB | ||
Filedesc metadata | emd-29039.cif.gz | 7.4 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-29039 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-29039 | HTTPS FTP |
-Validation report
Summary document | emd_29039_validation.pdf.gz | 401 KB | Display | EMDB validaton report |
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Full document | emd_29039_full_validation.pdf.gz | 400.5 KB | Display | |
Data in XML | emd_29039_validation.xml.gz | 7.4 KB | Display | |
Data in CIF | emd_29039_validation.cif.gz | 8.4 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-29039 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-29039 | HTTPS FTP |
-Related structure data
Related structure data | 8ff4MC 8fcjC 8fcuC 8fcvC 8fcwC 8fcxC 8fd2C 8fd3C 8ff5C C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_29039.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.054 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Sample components
+Entire : Structure of Cascade-DNA-TniQ-TnsC complex (composite) in type I-...
+Supramolecule #1: Structure of Cascade-DNA-TniQ-TnsC complex (composite) in type I-...
+Macromolecule #1: Type I-B CRISPR-associated protein Cas5
+Macromolecule #2: Type I-B CRISPR-associated protein Cas6
+Macromolecule #3: Type I-B CRISPR-associated protein Cas7
+Macromolecule #4: Type I-B CRISPR-associated protein Cas8
+Macromolecule #5: Type I-B CRISPR-associated protein Cas11
+Macromolecule #9: TniQ
+Macromolecule #10: TnsC
+Macromolecule #6: RNA
+Macromolecule #7: Target DNA strand
+Macromolecule #8: Non-target DNA strand
+Macromolecule #11: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #12: MAGNESIUM ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 54.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.8 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: OTHER / Details: Two startup models are used: 8FCU and 8FCV |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 281621 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |