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Yorodumi- EMDB-31332: The cryo-EM map of the DDX42-SF3b complex in N-terminus of DDX42 -
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Open data
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Basic information
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| Title | The cryo-EM map of the DDX42-SF3b complex in N-terminus of DDX42 | |||||||||
Map data | The EM map of DDX42 Nhelix region | |||||||||
Sample |
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Keywords | DDX32 / SF3B1 / SF3b / U2 snRNP / SPLICING | |||||||||
| Biological species | Homo sapiens (human) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 2.7 Å | |||||||||
Authors | Zhang X / Zhan X / Shi Y | |||||||||
| Funding support | China, 1 items
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Citation | Journal: Nat Commun / Year: 2023Title: Mechanisms of the RNA helicases DDX42 and DDX46 in human U2 snRNP assembly. Authors: Fenghua Yang / Tong Bian / Xiechao Zhan / Zhe Chen / Zhihan Xing / Nicolas A Larsen / Xiaofeng Zhang / Yigong Shi / ![]() Abstract: Three RNA helicases - DDX42, DDX46 and DHX15 - are found to be associated with human U2 snRNP, but their roles and mechanisms in U2 snRNP and spliceosome assembly are insufficiently understood. Here ...Three RNA helicases - DDX42, DDX46 and DHX15 - are found to be associated with human U2 snRNP, but their roles and mechanisms in U2 snRNP and spliceosome assembly are insufficiently understood. Here we report the cryo-electron microscopy (cryo-EM) structures of the DDX42-SF3b complex and a putative assembly precursor of 17S U2 snRNP that contains DDX42 (DDX42-U2 complex). DDX42 is anchored on SF3B1 through N-terminal sequences, with its N-plug occupying the RNA path of SF3B1. The binding mode of DDX42 to SF3B1 is in striking analogy to that of DDX46. In the DDX42-U2 complex, the N-terminus of DDX42 remains anchored on SF3B1, but the helicase domain has been displaced by U2 snRNA and TAT-SF1. Through in vitro assays, we show DDX42 and DDX46 are mutually exclusive in terms of binding to SF3b. Cancer-driving mutations of SF3B1 target the residues in the RNA path that directly interact with DDX42 and DDX46. These findings reveal the distinct roles of DDX42 and DDX46 in assembly of 17S U2 snRNP and provide insights into the mechanisms of SF3B1 cancer mutations. | |||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_31332.map.gz | 49.4 MB | EMDB map data format | |
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| Header (meta data) | emd-31332-v30.xml emd-31332.xml | 9 KB 9 KB | Display Display | EMDB header |
| Images | emd_31332.png | 91.2 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-31332 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-31332 | HTTPS FTP |
-Validation report
| Summary document | emd_31332_validation.pdf.gz | 577.5 KB | Display | EMDB validaton report |
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| Full document | emd_31332_full_validation.pdf.gz | 577.1 KB | Display | |
| Data in XML | emd_31332_validation.xml.gz | 5.9 KB | Display | |
| Data in CIF | emd_31332_validation.cif.gz | 6.7 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-31332 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-31332 | HTTPS FTP |
-Related structure data
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Map
| File | Download / File: emd_31332.map.gz / Format: CCP4 / Size: 52.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | The EM map of DDX42 Nhelix region | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.087 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 : The DDX42-SF3b core complex
| Entire | Name: The DDX42-SF3b core complex |
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| Components |
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-Supramolecule #1: The DDX42-SF3b core complex
| Supramolecule | Name: The DDX42-SF3b core complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#5 |
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| Source (natural) | Organism: Homo sapiens (human) |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 1.5 mg/mL |
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| Buffer | pH: 7.9 |
| Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD / Support film - Material: CARBON / Support film - topology: HOLEY |
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 50.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
| Startup model | Type of model: EMDB MAP |
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| Final reconstruction | Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 2.7 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 104710 |
| Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
| Final angle assignment | Type: MAXIMUM LIKELIHOOD |
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About Yorodumi



Keywords
Homo sapiens (human)
Authors
China, 1 items
Citation





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FIELD EMISSION GUN
