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Yorodumi- EMDB-9624: The Cryo-EM Structure of Human Pre-catalytic Spliceosome (B compl... -
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Basic information
| Entry | Database: EMDB / ID: EMD-9624 | |||||||||
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| Title | The Cryo-EM Structure of Human Pre-catalytic Spliceosome (B complex) at 3.8 angstrom resolution | |||||||||
Map data | The 3.8 Angstrom map of the human B complex | |||||||||
Sample |
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Keywords | Spliceosome / SPLICING | |||||||||
| Function / homology | Function and homology informationmicrofibril / 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 ...microfibril / 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 / U6 snRNP / U11/U12 snRNP / box C/D sno(s)RNA binding / PH domain binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / dense fibrillar component / cis assembly of pre-catalytic spliceosome / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / B-WICH complex / box C/D methylation guide snoRNP complex / protein methylation / U4/U6 snRNP / U12-type spliceosomal complex / 7-methylguanosine cap hypermethylation / U1 snRNP binding / methylosome / RNA splicing, via transesterification reactions / pICln-Sm protein complex / U2-type catalytic step 1 spliceosome / snRNP binding / blastocyst formation / small nuclear ribonucleoprotein complex / sno(s)RNA-containing ribonucleoprotein complex / splicing factor binding / SMN-Sm protein complex / proline-rich region binding / spliceosomal tri-snRNP complex / U4 snRNA binding / 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 / SAGA complex / RNA Polymerase II Transcription Termination / P-body assembly / U2 snRNP / U1 snRNP / positive regulation of protein targeting to mitochondrion / U4 snRNP / U2-type prespliceosome / rRNA modification in the nucleus and cytosol / U3 snoRNA binding / tRNA processing / positive regulation of transcription by RNA polymerase III / K63-linked polyubiquitin modification-dependent protein binding / cyclosporin A binding / precatalytic spliceosome / regulation of alternative mRNA splicing, via spliceosome / regulation of RNA splicing / mRNA catabolic process / spliceosomal complex assembly / mRNA Splicing - Minor Pathway / mRNA 3'-splice site recognition / positive regulation of transcription by RNA polymerase I / nuclear-transcribed mRNA catabolic process / MLL1 complex / spliceosomal tri-snRNP complex assembly / U5 snRNA binding / 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 / positive regulation of viral genome replication / RNA processing / U1 snRNA binding / Cajal body / regulation of DNA repair / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / Gene and protein expression by JAK-STAT signaling after Interleukin-12 stimulation / RNA splicing / response to cocaine / maturation of SSU-rRNA Similarity search - Function | |||||||||
| Biological species | Homo sapiens (human) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.8 Å | |||||||||
Authors | Zhan X / Yan C | |||||||||
| Funding support | China, 2 items
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Citation | Journal: Cell Res / Year: 2018Title: Structures of the human pre-catalytic spliceosome and its precursor spliceosome. Authors: Xiechao Zhan / Chuangye Yan / Xiaofeng Zhang / Jianlin Lei / Yigong Shi / ![]() Abstract: The pre-catalytic spliceosome (B complex) is preceded by its precursor spliceosome (pre-B complex) and followed by the activated spliceosome (B complex). The pre-B-to-B and B-to-B transitions are ...The pre-catalytic spliceosome (B complex) is preceded by its precursor spliceosome (pre-B complex) and followed by the activated spliceosome (B complex). The pre-B-to-B and B-to-B transitions are driven by the ATPase/helicases Prp28 and Brr2, respectively. In this study, we report the cryo-electron microscopy structures of the human pre-B complex and the human B complex at an average resolution of 5.7 and 3.8 Å, respectively. In the pre-B complex, U1 and U2 small nuclear ribonucleoproteins (snRNPs) associate with two edges of the tetrahedron-shaped U4/U6.U5 tri-snRNP. The pre-mRNA is yet to be recognized by U5 or U6 small nuclear RNA (snRNA), and loop I of U5 snRNA remains unengaged. In the B complex, U1 snRNP and Prp28 are dissociated, the 5'-exon is anchored to loop I of U5 snRNA, and the 5'-splice site is recognized by U6 snRNA through duplex formation. In sharp contrast to S. cerevisiae, most components of U2 snRNP and tri-snRNP, exemplified by Brr2, undergo pronounced rearrangements in the human pre-B-to-B transition. Structural analysis reveals mechanistic insights into the assembly and activation of the human spliceosome. | |||||||||
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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_9624.map.gz | 226.5 MB | EMDB map data format | |
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| Header (meta data) | emd-9624-v30.xml emd-9624.xml | 71.3 KB 71.3 KB | Display Display | EMDB header |
| Images | emd_9624.png | 98.5 KB | ||
| Filedesc metadata | emd-9624.cif.gz | 20.4 KB | ||
| Others | emd_9624_additional.map.gz | 227.5 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-9624 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-9624 | HTTPS FTP |
-Validation report
| Summary document | emd_9624_validation.pdf.gz | 611.4 KB | Display | EMDB validaton report |
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| Full document | emd_9624_full_validation.pdf.gz | 611 KB | Display | |
| Data in XML | emd_9624_validation.xml.gz | 7 KB | Display | |
| Data in CIF | emd_9624_validation.cif.gz | 8.1 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9624 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9624 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 6ahdMC ![]() 9621C ![]() 6ah0C 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_9624.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | The 3.8 Angstrom map of the human 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: The 4.5 Angstrom map of the human B complex
| File | emd_9624_additional.map | ||||||||||||
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| Annotation | The 4.5 Angstrom map of the human B complex | ||||||||||||
| Projections & Slices |
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| Density Histograms |
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Sample components
+Entire : Human Pre-catalytic spliceosome (B complex)
+Supramolecule #1: Human Pre-catalytic spliceosome (B complex)
+Macromolecule #1: Pre-mRNA-processing-splicing factor 8
+Macromolecule #6: Thioredoxin-like protein 4A
+Macromolecule #7: 116 kDa U5 small nuclear ribonucleoprotein component
+Macromolecule #8: Pre-mRNA-processing factor 6
+Macromolecule #9: NHP2-like protein 1
+Macromolecule #10: U4/U6 small nuclear ribonucleoprotein Prp31
+Macromolecule #11: U4/U6.U5 tri-snRNP-associated protein 1
+Macromolecule #12: U4/U6 small nuclear ribonucleoprotein Prp3
+Macromolecule #13: SmB
+Macromolecule #14: Small nuclear ribonucleoprotein Sm D1
+Macromolecule #15: Small nuclear ribonucleoprotein Sm D2
+Macromolecule #16: SmE
+Macromolecule #17: SmF
+Macromolecule #18: Small nuclear ribonucleoprotein G
+Macromolecule #19: Small nuclear ribonucleoprotein Sm D3
+Macromolecule #20: U5 small nuclear ribonucleoprotein 40 kDa protein
+Macromolecule #21: WW domain-binding protein 4
+Macromolecule #22: Peptidyl-prolyl cis-trans isomerase H
+Macromolecule #23: Ubiquitin-like protein 5
+Macromolecule #24: Microfibrillar-associated protein 1
+Macromolecule #25: Pre-mRNA-splicing factor 38A
+Macromolecule #26: Zinc finger matrin-type protein 2
+Macromolecule #27: WD40 repeat-containing protein SMU1
+Macromolecule #29: U2 small nuclear ribonucleoprotein A'
+Macromolecule #30: U2 small nuclear ribonucleoprotein B''
+Macromolecule #31: Splicing factor 3A subunit 1
+Macromolecule #32: Splicing factor 3A subunit 2
+Macromolecule #33: Splicing factor 3A subunit 3
+Macromolecule #34: U6 snRNA-associated Sm-like protein LSm2
+Macromolecule #35: U6 snRNA-associated Sm-like protein LSm3
+Macromolecule #36: U6 snRNA-associated Sm-like protein LSm4
+Macromolecule #37: U6 snRNA-associated Sm-like protein LSm5
+Macromolecule #38: U6 snRNA-associated Sm-like protein LSm6
+Macromolecule #39: U6 snRNA-associated Sm-like protein LSm7
+Macromolecule #40: U6 snRNA-associated Sm-like protein LSm8
+Macromolecule #41: U4/U6 small nuclear ribonucleoprotein Prp4
+Macromolecule #42: Splicing factor 3B subunit 1
+Macromolecule #43: Splicing factor 3B subunit 3
+Macromolecule #44: SF3b14a, Splicing factor 3B subunit 6
+Macromolecule #45: PHD finger-like domain-containing protein 5A
+Macromolecule #46: SF3b5, Splicing factor 3B subunit 5
+Macromolecule #47: SF3b145, Splicing factor 3B subunit 2
+Macromolecule #48: SF3b49, Splicing factor 3B subunit 4
+Macromolecule #49: Brr2, U5 small nuclear ribonucleoprotein 200 kDa helicase
+Macromolecule #2: U4snRNA
+Macromolecule #3: U5snRNA
+Macromolecule #4: U6snRNA
+Macromolecule #5: pre-mRNA
+Macromolecule #28: U2snRNA
+Macromolecule #50: INOSITOL HEXAKISPHOSPHATE
+Macromolecule #51: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #52: MAGNESIUM 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
| Buffer | pH: 7.9 |
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| 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: 45.0 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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Image processing
| Startup model | Type of model: PDB ENTRY |
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| Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.8 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 137853 |
| Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
| Final angle assignment | Type: MAXIMUM LIKELIHOOD |
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About Yorodumi


Keywords
Homo sapiens (human)
Authors
China, 2 items
Citation
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