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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-3979 | |||||||||
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| Title | Post-catalytic P complex spliceosome with 3' splice site docked | |||||||||
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Keywords | Spliceosome / P-complex / Exon ligation / SPLICING | |||||||||
| Function / homology | Function and homology informationU2-type post-spliceosomal complex / mRNA branch site recognition / spliceosomal complex disassembly / U2-type post-mRNA release spliceosomal complex / cellular bud site selection / pre-mRNA 3'-splice site binding / post-mRNA release spliceosomal complex / generation of catalytic spliceosome for first transesterification step / cis assembly of pre-catalytic spliceosome / nuclear mRNA surveillance ...U2-type post-spliceosomal complex / mRNA branch site recognition / spliceosomal complex disassembly / U2-type post-mRNA release spliceosomal complex / cellular bud site selection / pre-mRNA 3'-splice site binding / post-mRNA release spliceosomal complex / generation of catalytic spliceosome for first transesterification step / cis assembly of pre-catalytic spliceosome / nuclear mRNA surveillance / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U4/U6 snRNP / 7-methylguanosine cap hypermethylation / pICln-Sm protein complex / U2-type catalytic step 1 spliceosome / pre-mRNA binding / snRNP binding / small nuclear ribonucleoprotein complex / splicing factor binding / SMN-Sm protein complex / spliceosomal tri-snRNP complex / commitment complex / mRNA cis splicing, via spliceosome / U2-type spliceosomal complex / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / U2 snRNP / U1 snRNP / U4 snRNP / poly(U) RNA binding / U2-type prespliceosome / precatalytic spliceosome / generation of catalytic spliceosome for second transesterification step / Formation of TC-NER Pre-Incision Complex / mRNA 5'-splice site recognition / spliceosomal complex assembly / mRNA 3'-splice site recognition / Gap-filling DNA repair synthesis and ligation in TC-NER / DNA replication origin binding / spliceosomal tri-snRNP complex assembly / Prp19 complex / Dual incision in TC-NER / U5 snRNA binding / protein K63-linked ubiquitination / U5 snRNP / DNA replication initiation / U2 snRNA binding / U6 snRNA binding / pre-mRNA intronic binding / spliceosomal snRNP assembly / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / positive regulation of cell cycle / catalytic step 2 spliceosome / nuclear periphery / RNA splicing / positive regulation of RNA splicing / spliceosomal complex / mRNA splicing, via spliceosome / RING-type E3 ubiquitin transferase / metallopeptidase activity / ubiquitin-protein transferase activity / ubiquitin protein ligase activity / RNA helicase activity / RNA helicase / DNA repair / GTPase activity / mRNA binding / chromatin binding / chromatin / GTP binding / ATP hydrolysis activity / mitochondrion / DNA binding / RNA binding / zinc ion binding / ATP binding / metal ion binding / identical protein binding / nucleus / cytoplasm / cytosol Similarity search - Function | |||||||||
| Biological species | ![]() ![]() ![]() | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.7 Å | |||||||||
Authors | Wilkinson ME / Fica SM | |||||||||
| Funding support | United Kingdom, 2 items
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Citation | Journal: Science / Year: 2017Title: Postcatalytic spliceosome structure reveals mechanism of 3'-splice site selection. Authors: Max E Wilkinson / Sebastian M Fica / Wojciech P Galej / Christine M Norman / Andrew J Newman / Kiyoshi Nagai / ![]() Abstract: Introns are removed from eukaryotic messenger RNA precursors by the spliceosome in two transesterification reactions-branching and exon ligation. The mechanism of 3'-splice site recognition during ...Introns are removed from eukaryotic messenger RNA precursors by the spliceosome in two transesterification reactions-branching and exon ligation. The mechanism of 3'-splice site recognition during exon ligation has remained unclear. Here we present the 3.7-angstrom cryo-electron microscopy structure of the yeast P-complex spliceosome immediately after exon ligation. The 3'-splice site AG dinucleotide is recognized through non-Watson-Crick pairing with the 5' splice site and the branch-point adenosine. After the branching reaction, protein factors work together to remodel the spliceosome and stabilize a conformation competent for 3'-splice site docking, thereby promoting exon ligation. The structure accounts for the strict conservation of the GU and AG dinucleotides at the 5' and 3' ends of introns and provides insight into the catalytic mechanism of exon ligation. | |||||||||
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Structure visualization
| Movie |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_3979.map.gz | 390.9 MB | EMDB map data format | |
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| Header (meta data) | emd-3979-v30.xml emd-3979.xml | 74.9 KB 74.9 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_3979_fsc.xml | 17 KB | Display | FSC data file |
| Images | emd_3979.png | 71 KB | ||
| Filedesc metadata | emd-3979.cif.gz | 18.6 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-3979 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-3979 | HTTPS FTP |
-Validation report
| Summary document | emd_3979_validation.pdf.gz | 726.2 KB | Display | EMDB validaton report |
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| Full document | emd_3979_full_validation.pdf.gz | 725.7 KB | Display | |
| Data in XML | emd_3979_validation.xml.gz | 15.2 KB | Display | |
| Data in CIF | emd_3979_validation.cif.gz | 21.2 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-3979 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-3979 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 6exnMC ![]() 3980C M: atomic model generated by this map C: citing same article ( |
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| Similar structure data |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_3979.map.gz / Format: CCP4 / Size: 421.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.12 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 : P complex spliceosome with 3' exon docked
+Supramolecule #1: P complex spliceosome with 3' exon docked
+Supramolecule #2: P complex spliceosome
+Supramolecule #3: 3' exon
+Macromolecule #1: U2 snRNA
+Macromolecule #2: U5 snRNA
+Macromolecule #3: U6 snRNA
+Macromolecule #7: Ligated exons: UBC4 mRNA
+Macromolecule #9: Intron lariat: UBC4 RNA
+Macromolecule #4: Pre-mRNA-splicing factor Prp8
+Macromolecule #5: Pre-mRNA-splicing factor SNU114
+Macromolecule #6: Pre-mRNA-splicing factor CWC16
+Macromolecule #8: Pre-mRNA-splicing factor CWC22
+Macromolecule #10: Pre-mRNA-splicing factor PRP46
+Macromolecule #11: Pre-mRNA-processing protein 45
+Macromolecule #12: Pre-mRNA-splicing factor BUD31
+Macromolecule #13: Pre-mRNA-splicing factor CWC2
+Macromolecule #14: Pre-mRNA-splicing factor SLT11
+Macromolecule #15: Pre-mRNA-splicing factor CEF1
+Macromolecule #16: Pre-mRNA-splicing factor CWC15
+Macromolecule #17: Pre-mRNA-splicing factor CWC21
+Macromolecule #18: Pre-mRNA-splicing factor CLF1
+Macromolecule #19: Pre-mRNA-splicing factor SYF1
+Macromolecule #20: Pre-mRNA-splicing factor ATP-dependent RNA helicase PRP22
+Macromolecule #21: U2 small nuclear ribonucleoprotein A'
+Macromolecule #22: Unassigned structure
+Macromolecule #23: U2 small nuclear ribonucleoprotein B''
+Macromolecule #24: Pre-mRNA-splicing factor Prp18
+Macromolecule #25: Small nuclear ribonucleoprotein-associated protein B
+Macromolecule #26: Pre-mRNA-splicing factor SLU7
+Macromolecule #27: Small nuclear ribonucleoprotein Sm D3
+Macromolecule #28: Small nuclear ribonucleoprotein E
+Macromolecule #29: Small nuclear ribonucleoprotein F
+Macromolecule #30: Small nuclear ribonucleoprotein G
+Macromolecule #31: Small nuclear ribonucleoprotein Sm D1
+Macromolecule #32: Small nuclear ribonucleoprotein Sm D2
+Macromolecule #33: Pre-mRNA-processing factor Prp17
+Macromolecule #34: Pre-mRNA-splicing factor SNT309
+Macromolecule #35: Pre-mRNA-processing factor Prp19
+Macromolecule #36: Pre-mRNA-splicing factor SYF2
+Macromolecule #37: MAGNESIUM ION
+Macromolecule #38: INOSITOL HEXAKISPHOSPHATE
+Macromolecule #39: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #40: ZINC ION
-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 | 1.4 mg/mL | ||||||||||||
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| Buffer | pH: 7.9 Component:
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| Grid | Model: Quantifoil R2/2 / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 7 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 15 sec. / Pretreatment - Atmosphere: AIR | ||||||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK III Details: 3 uL sample was applied to the grid, left for 30s, then blotted for 3s and immediately plunged into liquid ethane.. |
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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: COUNTING / Digitization - Frames/image: 1-20 / Number grids imaged: 1 / Number real images: 2295 / Average exposure time: 12.0 sec. / Average electron dose: 47.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | C2 aperture diameter: 50.0 µm / Calibrated defocus max: 3.0 µm / Calibrated defocus min: 0.2 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal magnification: 105000 |
| 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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About Yorodumi



Keywords
Authors
United Kingdom, 2 items
Citation
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