+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-8014 | |||||||||
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Title | Body region of the U4/U6.U5 tri-snRNP | |||||||||
Map data | Central region of the U4/U6.U5 tri-snRNP comprising Prp8, Dib1, Prp31, Prp6, Prp4, Prp3, Snu13, U4 snRNA and U6 snRNA | |||||||||
Sample |
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Keywords | Transcription / snRNP / spliceosome / RNA-protein complex / U4/U6.U5 snRNP | |||||||||
Function / homology | Function and homology information spliceosomal conformational changes to generate catalytic conformation / snoRNA splicing / snoRNA guided rRNA 2'-O-methylation / box C/D sno(s)RNA 3'-end processing / box C/D methylation guide snoRNP complex / U4/U6 snRNP / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U4 snRNA binding / spliceosomal tri-snRNP complex / U3 snoRNA binding ...spliceosomal conformational changes to generate catalytic conformation / snoRNA splicing / snoRNA guided rRNA 2'-O-methylation / box C/D sno(s)RNA 3'-end processing / box C/D methylation guide snoRNP complex / U4/U6 snRNP / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U4 snRNA binding / spliceosomal tri-snRNP complex / U3 snoRNA binding / U4 snRNP / precatalytic spliceosome / Major pathway of rRNA processing in the nucleolus and cytosol / generation of catalytic spliceosome for second transesterification step / spliceosomal complex assembly / mRNA 5'-splice site recognition / mRNA 3'-splice site recognition / spliceosomal tri-snRNP complex assembly / U5 snRNA binding / U5 snRNP / U2 snRNA binding / U6 snRNA binding / spliceosomal snRNP assembly / pre-mRNA intronic binding / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / small-subunit processome / spliceosomal complex / mRNA splicing, via spliceosome / metallopeptidase activity / nucleic acid binding / RNA helicase activity / RNA helicase / mRNA binding / nucleolus / ATP hydrolysis activity / mitochondrion / RNA binding / nucleoplasm / ATP binding / identical protein binding / nucleus / cytoplasm Similarity search - Function | |||||||||
Biological species | Saccharomyces cerevisiae (brewer's yeast) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||
Authors | Nguyen THD / Galej WP | |||||||||
Funding support | United Kingdom, 1 items
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Citation | Journal: Nature / Year: 2016 Title: Cryo-EM structure of the yeast U4/U6.U5 tri-snRNP at 3.7 Å resolution. Authors: Thi Hoang Duong Nguyen / Wojciech P Galej / Xiao-Chen Bai / Chris Oubridge / Andrew J Newman / Sjors H W Scheres / Kiyoshi Nagai / Abstract: U4/U6.U5 tri-snRNP represents a substantial part of the spliceosome before activation. A cryo-electron microscopy structure of Saccharomyces cerevisiae U4/U6.U5 tri-snRNP at 3.7 Å resolution led ...U4/U6.U5 tri-snRNP represents a substantial part of the spliceosome before activation. A cryo-electron microscopy structure of Saccharomyces cerevisiae U4/U6.U5 tri-snRNP at 3.7 Å resolution led to an essentially complete atomic model comprising 30 proteins plus U4/U6 and U5 small nuclear RNAs (snRNAs). The structure reveals striking interweaving interactions of the protein and RNA components, including extended polypeptides penetrating into subunit interfaces. The invariant ACAGAGA sequence of U6 snRNA, which base-pairs with the 5'-splice site during catalytic activation, forms a hairpin stabilized by Dib1 and Prp8 while the adjacent nucleotides interact with the exon binding loop 1 of U5 snRNA. Snu114 harbours GTP, but its putative catalytic histidine is held away from the γ-phosphate by hydrogen bonding to a tyrosine in the amino-terminal domain of Prp8. Mutation of this histidine to alanine has no detectable effect on yeast growth. The structure provides important new insights into the spliceosome activation process leading to the formation of the catalytic centre. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_8014.map.gz | 196.2 MB | EMDB map data format | |
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Header (meta data) | emd-8014-v30.xml emd-8014.xml | 29.5 KB 29.5 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_8014_fsc.xml | 13.2 KB | Display | FSC data file |
Images | emd_8014.png | 233.6 KB | ||
Filedesc metadata | emd-8014.cif.gz | 11.2 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-8014 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-8014 | HTTPS FTP |
-Validation report
Summary document | emd_8014_validation.pdf.gz | 189.5 KB | Display | EMDB validaton report |
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Full document | emd_8014_full_validation.pdf.gz | 189.1 KB | Display | |
Data in XML | emd_8014_validation.xml.gz | 502 B | Display | |
Data in CIF | emd_8014_validation.cif.gz | 369 B | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8014 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8014 | HTTPS FTP |
-Related structure data
Related structure data | 5gapMC 8006C 8007C 8008C 8009C 8010C 8011C 8012C 8013C 5gamC 5ganC 5gaoC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_8014.map.gz / Format: CCP4 / Size: 209.3 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Central region of the U4/U6.U5 tri-snRNP comprising Prp8, Dib1, Prp31, Prp6, Prp4, Prp3, Snu13, U4 snRNA and U6 snRNA | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.43 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
+Entire : Body region of the U4/U6.U5 tri-snRNP complex
+Supramolecule #1: Body region of the U4/U6.U5 tri-snRNP complex
+Macromolecule #1: U4 snRNA, 5' region, nucleotides 1-67
+Macromolecule #2: U6 snRNA
+Macromolecule #3: U5 snRNA
+Macromolecule #4: unknown protein
+Macromolecule #5: Pre-mRNA-splicing factor 8
+Macromolecule #6: U4/U6 small nuclear ribonucleoprotein PRP4
+Macromolecule #7: Pre-mRNA-splicing factor 6
+Macromolecule #8: Spliceosomal protein DIB1
+Macromolecule #9: Pre-mRNA-processing factor 31
+Macromolecule #10: U4/U6 small nuclear ribonucleoprotein PRP3
+Macromolecule #11: 13 kDa ribonucleoprotein-associated protein
+Macromolecule #12: Pre-mRNA-splicing helicase BRR2
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.2 mg/mL |
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Buffer | pH: 7.9 / Component - Concentration: 1.0 mM / Component - Name: DTT |
Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 6 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 70 sec. / Pretreatment - Atmosphere: OTHER |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK III Details: Grids were blotted at 4 deg C for 2 seconds before plunging.. |
Details | 3.5 microlitre of sample was applied to grid. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Quantum |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Digitization - Frames/image: 1-20 / Number real images: 2477 / Average exposure time: 16.0 sec. / Average electron dose: 38.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Calibrated magnification: 35714 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.0 mm / Nominal defocus max: 3.5 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 81000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Refinement | Space: RECIPROCAL / Protocol: AB INITIO MODEL |
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Output model | PDB-5gap: |