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- EMDB-7818: Human nuclear exosome-MTR4 RNA complex - overall reconstruction -... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-7818 | |||||||||
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Title | Human nuclear exosome-MTR4 RNA complex - overall reconstruction - small class KOW domain open | |||||||||
![]() | Overall reconstruction for small class of human nuclear exosome-MTR4 complex with the MTR4 KOW domain in open conformation | |||||||||
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Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 7.75 Å | |||||||||
![]() | Weick E-M / Lima CD | |||||||||
![]() | ![]() Title: Helicase-Dependent RNA Decay Illuminated by a Cryo-EM Structure of a Human Nuclear RNA Exosome-MTR4 Complex. Authors: Eva-Maria Weick / M Rhyan Puno / Kurt Januszyk / John C Zinder / Michael A DiMattia / Christopher D Lima / ![]() Abstract: The ribonucleolytic RNA exosome interacts with RNA helicases to degrade RNA. To understand how the 3' to 5' Mtr4 helicase engages RNA and the nuclear exosome, we reconstituted 14-subunit Mtr4- ...The ribonucleolytic RNA exosome interacts with RNA helicases to degrade RNA. To understand how the 3' to 5' Mtr4 helicase engages RNA and the nuclear exosome, we reconstituted 14-subunit Mtr4-containing RNA exosomes from Saccharomyces cerevisiae, Schizosaccharomyces pombe, and human and show that they unwind structured substrates to promote degradation. We loaded a human exosome with an optimized DNA-RNA chimera that stalls MTR4 during unwinding and determined its structure to an overall resolution of 3.45 Å by cryoelectron microscopy (cryo-EM). The structure reveals an RNA-engaged helicase atop the non-catalytic core, with RNA captured within the central channel and DIS3 exoribonuclease active site. MPP6 tethers MTR4 to the exosome through contacts to the RecA domains of MTR4. EXOSC10 remains bound to the core, but its catalytic module and cofactor C1D are displaced by RNA-engaged MTR4. Competition for the exosome core may ensure that RNA is committed to degradation by DIS3 when engaged by MTR4. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 13.1 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 32.6 KB 32.6 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 13.8 KB | Display | ![]() |
Images | ![]() | 49.8 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 78 KB | Display | ![]() |
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Full document | ![]() | 77.1 KB | Display | |
Data in XML | ![]() | 494 B | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7808C ![]() 7809C ![]() 7810C ![]() 7812C ![]() 7813C ![]() 7814C ![]() 7815C ![]() 7819C ![]() 6d6qC ![]() 6d6rC C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Overall reconstruction for small class of human nuclear exosome-MTR4 complex with the MTR4 KOW domain in open conformation | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.07 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 : Human nuclear exosome-MTR4 helicase captured after unwinding a DN...
+Supramolecule #1: Human nuclear exosome-MTR4 helicase captured after unwinding a DN...
+Macromolecule #1: EXOSC9
+Macromolecule #2: EXOSC4
+Macromolecule #3: EXOSC8
+Macromolecule #4: EXOSC5
+Macromolecule #5: EXOSC7
+Macromolecule #6: EXOSC6
+Macromolecule #7: EXOSC3
+Macromolecule #8: EXOSC2
+Macromolecule #9: EXOSC1
+Macromolecule #10: EXOSC10
+Macromolecule #11: DIS3
+Macromolecule #12: MPP6, MPHOSPH6
+Macromolecule #13: MTR4, MTREX
+Macromolecule #14: C1D, Rrp47
+Macromolecule #15: synthetic RNA
+Macromolecule #16: DNA-RNA chimera
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 1.0 mg/mL | |||||||||||||||||||||
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Buffer | pH: 8 Component:
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Grid | Model: Quantifoil R2/2 / Material: GOLD / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 0.037 kPa | |||||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber temperature: 298 K / Instrument: FEI VITROBOT MARK IV / Details: 30 sec wait time, 2.5 sec blot time. | |||||||||||||||||||||
Details | Sample was monodisperse upon elution from gel filtration prior to vitrification. |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Number real images: 1439 / Average electron dose: 85.23 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Details | Models can be manually docked into this map. |
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