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- EMDB-10217: Cryo-EM structure of the RecBCD no Chi negative control complex -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-10217 | |||||||||
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Title | Cryo-EM structure of the RecBCD no Chi negative control complex | |||||||||
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![]() | RecBCD bound to a forked DNA substrate that does not contain Chi: RecBCD ![]() ![]() ![]() | |||||||||
Function / homology | ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() | |||||||||
Biological species | ![]() ![]() ![]() | |||||||||
Method | ![]() ![]() | |||||||||
![]() | Cheng K / Wilkinson M / Wigley DB | |||||||||
![]() | ![]() Title: A conformational switch in response to Chi converts RecBCD from phage destruction to DNA repair. Authors: Kaiying Cheng / Martin Wilkinson / Yuriy Chaban / Dale B Wigley / ![]() Abstract: The RecBCD complex plays key roles in phage DNA degradation, CRISPR array acquisition (adaptation) and host DNA repair. The switch between these roles is regulated by a DNA sequence called Chi. We ...The RecBCD complex plays key roles in phage DNA degradation, CRISPR array acquisition (adaptation) and host DNA repair. The switch between these roles is regulated by a DNA sequence called Chi. We report cryo-EM structures of the Escherichia coli RecBCD complex bound to several different DNA forks containing a Chi sequence, including one in which Chi is recognized and others in which it is not. The Chi-recognized structure shows conformational changes in regions of the protein that contact Chi and reveals a tortuous path taken by the DNA. Sequence specificity arises from interactions with both the RecC subunit and the sequence itself. These structures provide molecular details for how Chi is recognized and insights into the changes that occur in response to Chi binding that switch RecBCD from bacteriophage destruction and CRISPR spacer acquisition to constructive host DNA repair. | |||||||||
Validation Report | ![]() ![]() ![]() ![]() | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
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Download
FSC (resolution estimation) |
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Header (meta data in XML format) |
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Images |
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Archive directory |
-Related structure data
Related structure data | ![]() 6sjgCM ![]() 6sjbC ![]() 6sjeC ![]() 6sjfC ![]() 6t2uC ![]() 6t2vC C: citing same article ( M: atomic model generated by this map |
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Similar-shape strucutres |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.048 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
+Entire RecBCD bound to a forked DNA substrate that does not contain Chi
+Component #1: protein, RecBCD bound to a forked DNA substrate that does not con...
+Component #2: protein, RecBCD
+Component #3: protein, DNA
+Component #4: protein, RecBCD enzyme subunit RecB
+Component #5: protein, RecBCD enzyme subunit RecC
+Component #6: protein, RecBCD enzyme subunit RecD
+Component #7: nucleic-acid, Forked DNA substrate
-Experimental details
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Sample preparation
Specimen | Specimen state: Particle / Method: ![]() |
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Sample solution | Specimen conc.: 0.2 mg/mL / pH: 8 |
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Temperature: 277 K / Humidity: 90 % / Details: 1s blot before plunging. |
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Electron microscopy imaging
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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![]() | Microscope: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN![]() |
Lens | Magnification: 130000.0 X (nominal), 47710.0 X (calibrated) / Cs: 2.7 mm / Imaging mode: BRIGHT FIELD![]() |
Specimen Holder | Model: FEI TITAN KRIOS AUTOGRID HOLDER |
Camera | Detector: GATAN K2 SUMMIT (4k x 4k) |
-Image acquisition
Image acquisition | Number of digital images: 788 / Sampling size: 5 µm |
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