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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-4658 | |||||||||
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| Title | Structure of the human U5.U4/U6 tri-snRNP at 2.9A resolution. | |||||||||
Map data | Human U4.U5/U6 tri-snRNP, overall map. | |||||||||
Sample |
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Keywords | RNP complex / splicing / RNA / protein / spliceosome | |||||||||
| Function / homology | Function and homology informationLsm2-8 complex / U6 snRNA 3'-end binding / spliceosomal snRNP complex / ribonucleoprotein complex localization / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / RNA localization / U4atac snRNA binding / mRNA decay by 5' to 3' exoribonuclease ...Lsm2-8 complex / U6 snRNA 3'-end binding / spliceosomal snRNP complex / ribonucleoprotein complex localization / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / RNA localization / U4atac snRNA binding / mRNA decay by 5' to 3' exoribonuclease / Lsm1-7-Pat1 complex / R-loop processing / box C/D sno(s)RNA binding / U6 snRNP / PH domain binding / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / dense fibrillar component / histone pre-mRNA 3'end processing complex / cis assembly of pre-catalytic spliceosome / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / box C/D methylation guide snoRNP complex / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U4/U6 snRNP / protein methylation / U12-type spliceosomal complex / 7-methylguanosine cap hypermethylation / U1 snRNP binding / methylosome / sno(s)RNA-containing ribonucleoprotein complex / RNA splicing, via transesterification reactions / pICln-Sm protein complex / U2-type catalytic step 1 spliceosome / snRNP binding / U4 snRNA binding / small nuclear ribonucleoprotein complex / SMN-Sm protein complex / spliceosomal tri-snRNP complex / P granule / telomerase holoenzyme complex / U2-type precatalytic spliceosome / commitment complex / mRNA cis splicing, via spliceosome / telomerase RNA binding / U2-type spliceosomal complex / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / box C/D snoRNP assembly / U4 snRNP / RNA Polymerase II Transcription Termination / P-body assembly / U2 snRNP / U1 snRNP / U3 snoRNA binding / U2-type prespliceosome / rRNA modification in the nucleus and cytosol / tRNA processing / K63-linked polyubiquitin modification-dependent protein binding / precatalytic spliceosome / spliceosomal complex assembly / mRNA catabolic process / mRNA Splicing - Minor Pathway / negative regulation of mRNA splicing, via spliceosome / nuclear-transcribed mRNA catabolic process / MLL1 complex / spliceosomal tri-snRNP complex assembly / U5 snRNA binding / protein deubiquitination / U5 snRNP / U2 snRNA binding / U6 snRNA binding / pre-mRNA intronic binding / single fertilization / Major pathway of rRNA processing in the nucleolus and cytosol / spliceosomal snRNP assembly / ribonucleoprotein complex binding / RNA processing / U1 snRNA binding / Cajal body / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / RNA splicing / maturation of SSU-rRNA / response to cocaine / response to bacterium / spliceosomal complex / small-subunit processome / P-body / helicase activity / mRNA splicing, via spliceosome / small GTPase binding / cellular response to xenobiotic stimulus / mRNA processing / osteoblast differentiation / cellular response to tumor necrosis factor / cellular response to lipopolysaccharide Similarity search - Function | |||||||||
| Biological species | Homo sapiens (human) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 2.92 Å | |||||||||
Authors | Charenton C / Wilkinson ME | |||||||||
| Funding support | United Kingdom, 2 items
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Citation | Journal: Science / Year: 2019Title: Mechanism of 5' splice site transfer for human spliceosome activation. Authors: Clément Charenton / Max E Wilkinson / Kiyoshi Nagai / ![]() Abstract: The prespliceosome, comprising U1 and U2 small nuclear ribonucleoproteins (snRNPs) bound to the precursor messenger RNA 5' splice site (5'SS) and branch point sequence, associates with the U4/U6.U5 ...The prespliceosome, comprising U1 and U2 small nuclear ribonucleoproteins (snRNPs) bound to the precursor messenger RNA 5' splice site (5'SS) and branch point sequence, associates with the U4/U6.U5 tri-snRNP to form the fully assembled precatalytic pre-B spliceosome. Here, we report cryo-electron microscopy structures of the human pre-B complex captured before U1 snRNP dissociation at 3.3-angstrom core resolution and the human tri-snRNP at 2.9-angstrom resolution. U1 snRNP inserts the 5'SS-U1 snRNA helix between the two RecA domains of the Prp28 DEAD-box helicase. Adenosine 5'-triphosphate-dependent closure of the Prp28 RecA domains releases the 5'SS to pair with the nearby U6 ACAGAGA-box sequence presented as a mobile loop. The structures suggest that formation of the 5'SS-ACAGAGA helix triggers remodeling of an intricate protein-RNA network to induce Brr2 helicase relocation to its loading sequence in U4 snRNA, enabling Brr2 to unwind the U4/U6 snRNA duplex to allow U6 snRNA to form the catalytic center of the spliceosome. | |||||||||
| History |
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Structure visualization
| Movie |
Movie viewer |
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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_4658.map.gz | 260.2 MB | EMDB map data format | |
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| Header (meta data) | emd-4658-v30.xml emd-4658.xml | 60.5 KB 60.5 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_4658_fsc.xml | 14.8 KB | Display | FSC data file |
| Images | emd_4658.png | 181.4 KB | ||
| Filedesc metadata | emd-4658.cif.gz | 16.6 KB | ||
| Others | emd_4658_additional.map.gz | 260.1 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-4658 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-4658 | HTTPS FTP |
-Validation report
| Summary document | emd_4658_validation.pdf.gz | 624.6 KB | Display | EMDB validaton report |
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| Full document | emd_4658_full_validation.pdf.gz | 624.2 KB | Display | |
| Data in XML | emd_4658_validation.xml.gz | 14 KB | Display | |
| Data in CIF | emd_4658_validation.cif.gz | 19.3 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4658 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4658 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 6qw6MC ![]() 4665C ![]() 4672C ![]() 4673C ![]() 4674C ![]() 4675C ![]() 4676C ![]() 4686C ![]() 4687C ![]() 4688C ![]() 4689C ![]() 4690C ![]() 6qx9C M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | |
| EM raw data | EMPIAR-10307 (Title: Human pre-B spliceosome and U4/U6.U5 tri-snRNP / Data size: 2.3 TBData #1: Dataset 1 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #2: Dataset 2 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #3: Dataset 3 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #4: Dataset 4 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #5: Dataset 5 of human pre-B spliceosome; motion-corrected micrographs [micrographs - single frame] Data #6: Selected U4/U6.U5 tri-snRNP particles after Bayesian polishing [picked particles - single frame - processed] Data #7: Crude shifted preB particles [picked particles - single frame - unprocessed]) |
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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_4658.map.gz / Format: CCP4 / Size: 282.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | Human U4.U5/U6 tri-snRNP, overall map. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.022 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: Composite map from the multi body refinement. For...
| File | emd_4658_additional.map | ||||||||||||
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| Annotation | Composite map from the multi body refinement. For each map voxel, the map value was taken from the body with the highest local resolution as calculated with RELION. | ||||||||||||
| Projections & Slices |
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| Density Histograms |
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Sample components
+Entire : Human U4/U6.U5 tri-snRNP
+Supramolecule #1: Human U4/U6.U5 tri-snRNP
+Supramolecule #2: U5.U4/U6 tri-snRNP
+Supramolecule #3: Probable ATP-dependent RNA helicase DDX23
+Macromolecule #1: U4/U6.U5 small nuclear ribonucleoprotein 27 kDa protein
+Macromolecule #3: Small nuclear ribonucleoprotein Sm D1
+Macromolecule #4: Small nuclear ribonucleoprotein Sm D2
+Macromolecule #5: Small nuclear ribonucleoprotein Sm D3
+Macromolecule #6: U4/U6 small nuclear ribonucleoprotein Prp3
+Macromolecule #7: U4/U6 small nuclear ribonucleoprotein Prp4
+Macromolecule #8: U4/U6 small nuclear ribonucleoprotein Prp31
+Macromolecule #9: NHP2-like protein 1
+Macromolecule #10: Small nuclear ribonucleoprotein-associated proteins B and B'
+Macromolecule #11: Small nuclear ribonucleoprotein E
+Macromolecule #12: Small nuclear ribonucleoprotein F
+Macromolecule #13: Small nuclear ribonucleoprotein G
+Macromolecule #15: Pre-mRNA-processing-splicing factor 8
+Macromolecule #16: U5 small nuclear ribonucleoprotein 200 kDa helicase
+Macromolecule #17: 116 kDa U5 small nuclear ribonucleoprotein component
+Macromolecule #18: Thioredoxin-like protein 4A
+Macromolecule #19: Pre-mRNA-processing factor 6
+Macromolecule #20: U5 small nuclear ribonucleoprotein 40 kDa protein
+Macromolecule #21: Probable ATP-dependent RNA helicase DDX23
+Macromolecule #23: U6 snRNA-associated Sm-like protein LSm2
+Macromolecule #24: U6 snRNA-associated Sm-like protein LSm3
+Macromolecule #25: U6 snRNA-associated Sm-like protein LSm4
+Macromolecule #26: U6 snRNA-associated Sm-like protein LSm5
+Macromolecule #27: U6 snRNA-associated Sm-like protein LSm6
+Macromolecule #28: U6 snRNA-associated Sm-like protein LSm7
+Macromolecule #29: U6 snRNA-associated Sm-like protein LSm8
+Macromolecule #30: RNA-binding protein 42
+Macromolecule #31: U4/U6.U5 tri-snRNP-associated protein 1
+Macromolecule #32: U4/U6.U5 tri-snRNP-associated protein 2
+Macromolecule #2: U4 snRNA
+Macromolecule #14: U5 snRNA
+Macromolecule #22: U6 snRNA
+Macromolecule #33: INOSITOL HEXAKISPHOSPHATE
+Macromolecule #34: MAGNESIUM ION
+Macromolecule #35: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #36: 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 | 1.3 mg/mL | ||||||||
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| Buffer | pH: 7.9 Component:
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| Grid | Model: Quantifoil R2/2 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 6 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 20 sec. | ||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK III / Details: Wait 30s, blot for 2s to 3s.. |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Frames/image: 1-40 / Average exposure time: 6.0 sec. / Average electron dose: 50.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal magnification: 130000 |
| 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
Homo sapiens (human)
Authors
United Kingdom, 2 items
Citation
UCSF Chimera









































Z (Sec.)
Y (Row.)
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