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Yorodumi- EMDB-6974: Cryo-EM structure of the yeast B complex at average resolution of... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-6974 | ||||||||||||
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Title | Cryo-EM structure of the yeast B complex at average resolution of 3.9 angstrom | ||||||||||||
Map data | cryo-EM structure of the spliceosomal B complex | ||||||||||||
Sample |
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Keywords | spliceosome / assembly / B complex / SPLICING | ||||||||||||
Function / homology | Function and homology information maintenance of RNA location / spliceosomal conformational changes to generate catalytic conformation / RES complex / mRNA decay by 5' to 3' exoribonuclease / snoRNA splicing / maturation of 5S rRNA / snoRNA guided rRNA 2'-O-methylation / Lsm1-7-Pat1 complex / U6 snRNP / box C/D sno(s)RNA 3'-end processing ...maintenance of RNA location / spliceosomal conformational changes to generate catalytic conformation / RES complex / mRNA decay by 5' to 3' exoribonuclease / snoRNA splicing / maturation of 5S rRNA / snoRNA guided rRNA 2'-O-methylation / Lsm1-7-Pat1 complex / U6 snRNP / box C/D sno(s)RNA 3'-end processing / deadenylation-dependent decapping of nuclear-transcribed mRNA / generation of catalytic spliceosome for first transesterification step / box C/D methylation guide snoRNP complex / splicing factor binding / U4/U6 snRNP / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / 7-methylguanosine cap hypermethylation / pICln-Sm protein complex / P-body assembly / sno(s)RNA-containing ribonucleoprotein complex / snRNP binding / small nuclear ribonucleoprotein complex / U4 snRNA binding / SMN-Sm protein complex / spliceosomal tri-snRNP complex / U2-type spliceosomal complex / mRNA cis splicing, via spliceosome / poly(U) RNA binding / U3 snoRNA binding / commitment complex / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / U4 snRNP / U2 snRNP / U1 snRNP / U2-type prespliceosome / tRNA processing / 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 / nuclear-transcribed mRNA catabolic process / mRNA 3'-splice site recognition / spliceosomal tri-snRNP complex assembly / U5 snRNA binding / U5 snRNP / mRNA export from nucleus / U2 snRNA binding / U6 snRNA binding / spliceosomal snRNP assembly / cellular response to glucose starvation / 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 / P-body / mRNA splicing, via spliceosome / rRNA processing / metallopeptidase activity / nucleic acid binding / RNA helicase activity / RNA helicase / response to xenobiotic stimulus / GTPase activity / mRNA binding / GTP binding / nucleolus / ATP hydrolysis activity / mitochondrion / RNA binding / zinc ion binding / nucleoplasm / ATP binding / identical protein binding / nucleus / metal ion binding / cytosol / cytoplasm Similarity search - Function | ||||||||||||
Biological species | Saccharomyces cerevisiae S288c (yeast) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.9 Å | ||||||||||||
Authors | Shi Y / Bai R | ||||||||||||
Funding support | China, 3 items
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Citation | Journal: Science / Year: 2018 Title: Structures of the fully assembled spliceosome before activation. Authors: Rui Bai / Ruixue Wan / Chuangye Yan / Jianlin Lei / Yigong Shi / Abstract: The precatalytic spliceosome (B complex) is preceded by the pre-B complex. Here we report the cryo-electron microscopy structures of the pre-B and B complexes at average resolutions of 3.3 to 4.6 ...The precatalytic spliceosome (B complex) is preceded by the pre-B complex. Here we report the cryo-electron microscopy structures of the pre-B and B complexes at average resolutions of 3.3 to 4.6 and 3.9 angstroms, respectively. In the pre-B complex, the duplex between the 5' splice site (5'SS) and U1 small nuclear RNA (snRNA) is recognized by Yhc1, Luc7, and the Sm ring. In the B complex, U1 small nuclear ribonucleoprotein is dissociated, the 5'-exon-5'SS sequences are translocated near U6 snRNA, and three B-specific proteins may orient the precursor messenger RNA. In both complexes, U6 snRNA is anchored to loop I of U5 snRNA, and the duplex between the branch point sequence and U2 snRNA is recognized by the SF3b complex. Structural analysis reveals the mechanism of assembly and activation for the yeast spliceosome. | ||||||||||||
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_6974.map.gz | 227.3 MB | EMDB map data format | |
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Header (meta data) | emd-6974-v30.xml emd-6974.xml | 69.3 KB 69.3 KB | Display Display | EMDB header |
Images | emd_6974.png | 170.7 KB | ||
Filedesc metadata | emd-6974.cif.gz | 18.6 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-6974 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-6974 | HTTPS FTP |
-Validation report
Summary document | emd_6974_validation.pdf.gz | 560.9 KB | Display | EMDB validaton report |
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Full document | emd_6974_full_validation.pdf.gz | 560.5 KB | Display | |
Data in XML | emd_6974_validation.xml.gz | 7.1 KB | Display | |
Data in CIF | emd_6974_validation.cif.gz | 8.1 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-6974 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-6974 | HTTPS FTP |
-Related structure data
Related structure data | 5zwoMC 6972C 6973C 5zwmC 5zwnC C: citing same article (ref.) M: atomic model generated by this map |
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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_6974.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | cryo-EM structure of the spliceosomal B complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.338 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 : yeast fully assembled spliceosomal B complex before activation
+Supramolecule #1: yeast fully assembled spliceosomal B complex before activation
+Macromolecule #1: Pre-mRNA-splicing factor 8
+Macromolecule #2: U4/U6 small nuclear ribonucleoprotein PRP4
+Macromolecule #3: Pre-mRNA-processing factor 31
+Macromolecule #4: Pre-mRNA-splicing factor 6
+Macromolecule #5: U4/U6 small nuclear ribonucleoprotein PRP3
+Macromolecule #6: Spliceosomal protein DIB1
+Macromolecule #7: 13 kDa ribonucleoprotein-associated protein
+Macromolecule #8: Pre-mRNA-splicing factor SNU114
+Macromolecule #9: U6 snRNA-associated Sm-like protein LSm8
+Macromolecule #10: U6 snRNA-associated Sm-like protein LSm2
+Macromolecule #11: U6 snRNA-associated Sm-like protein LSm3
+Macromolecule #12: U6 snRNA-associated Sm-like protein LSm6
+Macromolecule #13: U6 snRNA-associated Sm-like protein LSm5
+Macromolecule #14: U6 snRNA-associated Sm-like protein LSm7
+Macromolecule #15: U6 snRNA-associated Sm-like protein LSm4
+Macromolecule #18: 66 kDa U4/U6.U5 small nuclear ribonucleoprotein component
+Macromolecule #19: Small nuclear ribonucleoprotein Sm D3
+Macromolecule #20: Small nuclear ribonucleoprotein-associated protein B
+Macromolecule #21: Small nuclear ribonucleoprotein Sm D1
+Macromolecule #22: Small nuclear ribonucleoprotein Sm D2
+Macromolecule #23: Small nuclear ribonucleoprotein E
+Macromolecule #24: Small nuclear ribonucleoprotein F
+Macromolecule #25: Small nuclear ribonucleoprotein G
+Macromolecule #27: Pre-mRNA-splicing helicase BRR2
+Macromolecule #29: U2 snRNP component HSH155
+Macromolecule #30: Cold sensitive U2 snRNA suppressor 1
+Macromolecule #31: Pre-mRNA-splicing factor RSE1
+Macromolecule #32: Protein HSH49
+Macromolecule #33: Pre-mRNA-splicing factor RDS3
+Macromolecule #34: RDS3 complex subunit 10
+Macromolecule #35: U2 snRNP component IST3
+Macromolecule #36: Pre-mRNA-splicing factor CWC26
+Macromolecule #37: Pre-mRNA leakage protein 1
+Macromolecule #39: U2 small nuclear ribonucleoprotein A'
+Macromolecule #40: U2 small nuclear ribonucleoprotein B''
+Macromolecule #41: Pre-mRNA-splicing factor PRP9
+Macromolecule #42: Pre-mRNA-splicing factor PRP21
+Macromolecule #43: Pre-mRNA-splicing factor PRP11
+Macromolecule #44: 23 kDa U4/U6.U5 small nuclear ribonucleoprotein component
+Macromolecule #45: Pre-mRNA-splicing factor 38
+Macromolecule #46: Pre-mRNA-splicing factor SPP381
+Macromolecule #16: U6 snRNA
+Macromolecule #17: U5 snRNA
+Macromolecule #26: U4 snRNA
+Macromolecule #28: Pre-mRNA
+Macromolecule #38: U2 snRNA
+Macromolecule #47: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #48: MAGNESIUM ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | cell |
-Sample preparation
Buffer | pH: 7.9 |
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Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k 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 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: EMDB MAP EMDB ID: |
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Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 2.0) / Number images used: 342588 |
Initial angle assignment | Type: PROJECTION MATCHING / Software - Name: RELION (ver. 2.0) |
Final angle assignment | Type: PROJECTION MATCHING / Software - Name: RELION (ver. 2.0) |