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Yorodumi- EMDB-6972: Cryo-EM structure of the yeast pre-B complex at an average resolu... -
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Basic information
| Entry | Database: EMDB / ID: EMD-6972 | ||||||||||||
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| Title | Cryo-EM structure of the yeast pre-B complex at an average resolution of 3.4~4.6 angstrom (tri-snRNP and U2 snRNP Part) | ||||||||||||
 Map data | tri-snRNP region of spliceosomal pre-B complex | ||||||||||||
 Sample | 
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 Keywords | spliceosome / assembly / pre-B complex / U1 snRNP / SPLICING | ||||||||||||
| Function / homology |  Function and homology informationmaintenance of RNA location / RES complex / spliceosomal conformational changes to generate catalytic conformation / 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 / deadenylation-dependent decapping of nuclear-transcribed mRNA ...maintenance of RNA location / RES complex / spliceosomal conformational changes to generate catalytic conformation / 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 / deadenylation-dependent decapping of nuclear-transcribed mRNA / box C/D sno(s)RNA 3'-end processing / generation of catalytic spliceosome for first transesterification step / box C/D methylation guide snoRNP complex / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U4/U6 snRNP / 7-methylguanosine cap hypermethylation / sno(s)RNA-containing ribonucleoprotein complex / pICln-Sm protein complex / snRNP binding / U4 snRNA 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 / U4 snRNP / P-body assembly / U2 snRNP / U1 snRNP / poly(U) RNA binding / U3 snoRNA binding / U2-type prespliceosome / tRNA processing / precatalytic spliceosome / generation of catalytic spliceosome for second transesterification step / Major pathway of rRNA processing in the nucleolus and cytosol / spliceosomal complex assembly / mRNA 5'-splice site recognition / mRNA 3'-splice site recognition / nuclear-transcribed mRNA catabolic process / spliceosomal tri-snRNP complex assembly / Prp19 complex / U5 snRNA binding / U5 snRNP / U2 snRNA binding / U6 snRNA binding / pre-mRNA intronic binding / spliceosomal snRNP assembly / mRNA export from nucleus / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / cellular response to glucose starvation / catalytic step 2 spliceosome / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / maturation of SSU-rRNA / spliceosomal complex / small-subunit processome / P-body / mRNA splicing, via spliceosome / metallopeptidase activity / rRNA processing / 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 / cytoplasm / cytosol Similarity search - Function  | ||||||||||||
| Biological species | ![]() ![]()  | ||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.4 Å | ||||||||||||
 Authors | Bai R / Wan R | ||||||||||||
| Funding support |   China, 3 items 
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 Citation |  Journal: Science / Year: 2018Title: 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 | 
 
 
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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_6972.map.gz | 226.7 MB |  EMDB map data format | |
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| Header (meta data) |  emd-6972-v30.xml emd-6972.xml | 72.2 KB 72.2 KB  | Display Display  |  EMDB header | 
| Images |  emd_6972.png | 228.4 KB | ||
| Filedesc metadata |  emd-6972.cif.gz | 18.3 KB | ||
| Others |  emd_6972_additional_1.map.gz emd_6972_additional_2.map.gz emd_6972_additional_3.map.gz | 222.8 MB 218.7 MB 226.8 MB  | ||
| Archive directory |  http://ftp.pdbj.org/pub/emdb/structures/EMD-6972 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-6972 | HTTPS FTP  | 
-Validation report
| Summary document |  emd_6972_validation.pdf.gz | 605.5 KB | Display |  EMDB validaton report | 
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| Full document |  emd_6972_full_validation.pdf.gz | 605.1 KB | Display | |
| Data in XML |  emd_6972_validation.xml.gz | 6.9 KB | Display | |
| Data in CIF |  emd_6972_validation.cif.gz | 8 KB | Display | |
| Arichive directory |  https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-6972 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-6972 | HTTPS FTP  | 
-Related structure data
| Related structure data | ![]() 5zwmMC ![]() 6973C ![]() 6974C ![]() 5zwnC ![]() 5zwoC C: citing same article ( M: atomic model generated by this map  | 
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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_6972.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | tri-snRNP region of spliceosomal pre-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
-Additional map: #3
| File | emd_6972_additional_1.map | ||||||||||||
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| Density Histograms | 
-Additional map: #2
| File | emd_6972_additional_2.map | ||||||||||||
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| Projections & Slices | 
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| Density Histograms | 
-Additional map: #1
| File | emd_6972_additional_3.map | ||||||||||||
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| Projections & Slices | 
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| Density Histograms | 
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Sample components
+Entire : yeast fully assembled spliceosomal pre-B complex before activation
+Supramolecule #1: yeast fully assembled spliceosomal pre-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 #19: 66 kDa U4/U6.U5 small nuclear ribonucleoprotein component
+Macromolecule #20: Small nuclear ribonucleoprotein Sm D3
+Macromolecule #21: Small nuclear ribonucleoprotein-associated protein B
+Macromolecule #22: Small nuclear ribonucleoprotein Sm D1
+Macromolecule #23: Small nuclear ribonucleoprotein Sm D2
+Macromolecule #24: Small nuclear ribonucleoprotein E
+Macromolecule #25: Small nuclear ribonucleoprotein F
+Macromolecule #26: Small nuclear ribonucleoprotein G
+Macromolecule #27: Pre-mRNA-splicing helicase BRR2
+Macromolecule #30: U2 small nuclear ribonucleoprotein A'
+Macromolecule #31: U2 small nuclear ribonucleoprotein B''
+Macromolecule #32: U2 snRNP component HSH155
+Macromolecule #33: Cold sensitive U2 snRNA suppressor 1
+Macromolecule #34: Pre-mRNA-splicing factor RSE1
+Macromolecule #35: Protein HSH49
+Macromolecule #36: Pre-mRNA-splicing factor RDS3
+Macromolecule #37: RDS3 complex subunit 10
+Macromolecule #38: U2 snRNP component IST3
+Macromolecule #39: Pre-mRNA-splicing factor CWC26
+Macromolecule #40: Pre-mRNA leakage protein 1
+Macromolecule #41: Pre-mRNA-splicing factor PRP9
+Macromolecule #42: Pre-mRNA-splicing factor PRP21
+Macromolecule #43: Pre-mRNA-splicing factor PRP11
+Macromolecule #16: U6 snRNA
+Macromolecule #17: U4 snRNA
+Macromolecule #18: U5 snRNA
+Macromolecule #28: Pre-mRNA-BPS
+Macromolecule #29: U2 snRNA
+Macromolecule #44: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #45: MAGNESIUM ION
+Macromolecule #46: ZINC ION
-Experimental details
-Structure determination
| Method | cryo EM | 
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 Processing | single particle reconstruction | 
| Aggregation state | cell | 
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Sample preparation
| Buffer | pH: 7.9 | 
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| Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER | 
| Vitrification | Cryogen name: 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) / 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  | 
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Keywords
Authors
China, 3 items 
Citation
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