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- EMDB-29039: Cryo-EM structure of Cascade-DNA-TniQ-TnsC complex (composite) in... -
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Open data
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Basic information
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Title | Cryo-EM structure of Cascade-DNA-TniQ-TnsC complex (composite) in type I-B CAST system | |||||||||
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![]() | CRISPR / transposon / TnsC / TniQ / DNA BINDING PROTEIN | |||||||||
Function / homology | ![]() | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||
![]() | Chang L / Wang S | |||||||||
Funding support | ![]()
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![]() | ![]() 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 | ![]() | 16 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 23.4 KB 23.4 KB | Display Display | ![]() |
Images | ![]() | 116 KB | ||
Filedesc metadata | ![]() | 7.4 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 401 KB | Display | ![]() |
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Full document | ![]() | 400.5 KB | Display | |
Data in XML | ![]() | 7.4 KB | Display | |
Data in CIF | ![]() | 8.4 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8ff4MC ![]() 8fcjC ![]() 8fcuC ![]() 8fcvC ![]() 8fcwC ![]() 8fcxC ![]() 8fd2C ![]() 8fd3C ![]() 8ff5C C: citing same article ( M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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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
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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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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: ETHANE |
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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: ![]() |
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 |
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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 |