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Yorodumi- EMDB-9525: Cryo-EM structure of the Catalytic Step I spliceosome (C complex)... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-9525 | |||||||||
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Title | Cryo-EM structure of the Catalytic Step I spliceosome (C complex) at 3.4 angstrom resolution | |||||||||
Map data | Cryo-EM map of Catalytic Step I spliceosome (C complex) at 3.4 angstrom | |||||||||
Sample |
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Keywords | RNA splicing / Spliceosome / Catalytic Step I / Intron lariat / RNA BINDING PROTEIN-RNA complex | |||||||||
Function / homology | Function and homology information post-spliceosomal complex / mRNA branch site recognition / U2-type post-mRNA release spliceosomal complex / cellular bud site selection / post-mRNA release spliceosomal complex / generation of catalytic spliceosome for first transesterification step / cis assembly of pre-catalytic spliceosome / splicing factor binding / U4/U6 snRNP / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) ...post-spliceosomal complex / mRNA branch site recognition / U2-type post-mRNA release spliceosomal complex / cellular bud site selection / post-mRNA release spliceosomal complex / generation of catalytic spliceosome for first transesterification step / cis assembly of pre-catalytic spliceosome / splicing factor binding / U4/U6 snRNP / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / 7-methylguanosine cap hypermethylation / Prp19 complex / pICln-Sm protein complex / snRNP binding / U2-type catalytic step 1 spliceosome / small nuclear ribonucleoprotein complex / pre-mRNA binding / SMN-Sm protein complex / spliceosomal tri-snRNP complex / U2-type spliceosomal complex / mRNA cis splicing, via spliceosome / poly(U) RNA binding / commitment complex / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / U4 snRNP / U2 snRNP / U1 snRNP / U2-type prespliceosome / precatalytic spliceosome / Formation of TC-NER Pre-Incision Complex / generation of catalytic spliceosome for second transesterification step / spliceosomal complex assembly / DNA replication origin binding / mRNA 5'-splice site recognition / Gap-filling DNA repair synthesis and ligation in TC-NER / protein K63-linked ubiquitination / mRNA 3'-splice site recognition / Dual incision in TC-NER / spliceosomal tri-snRNP complex assembly / DNA replication initiation / U5 snRNA binding / positive regulation of cell cycle / U5 snRNP / U2 snRNA binding / U6 snRNA binding / spliceosomal snRNP assembly / pre-mRNA intronic binding / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / RNA splicing / nuclear periphery / positive regulation of RNA splicing / spliceosomal complex / RING-type E3 ubiquitin transferase / mRNA splicing, via spliceosome / ubiquitin-protein transferase activity / metallopeptidase activity / ubiquitin protein ligase activity / DNA repair / GTPase activity / mRNA binding / chromatin binding / chromatin / GTP binding / mitochondrion / DNA binding / RNA binding / identical protein binding / nucleus / metal ion binding / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Saccharomyces cerevisiae S288c (yeast) / Saccharomyces cerevisiae S288C (yeast) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.4 Å | |||||||||
Authors | Wan R / Yan C | |||||||||
Citation | Journal: Science / Year: 2016 Title: Structure of a yeast catalytic step I spliceosome at 3.4 Å resolution. Authors: Ruixue Wan / Chuangye Yan / Rui Bai / Gaoxingyu Huang / Yigong Shi / Abstract: Each cycle of pre-messenger RNA splicing, carried out by the spliceosome, comprises two sequential transesterification reactions, which result in the removal of an intron and the joining of two exons. ...Each cycle of pre-messenger RNA splicing, carried out by the spliceosome, comprises two sequential transesterification reactions, which result in the removal of an intron and the joining of two exons. Here we report an atomic structure of a catalytic step I spliceosome (known as the C complex) from Saccharomyces cerevisiae, as determined by cryo-electron microscopy at an average resolution of 3.4 angstroms. In the structure, the 2'-OH of the invariant adenine nucleotide in the branch point sequence (BPS) is covalently joined to the phosphate at the 5' end of the 5' splice site (5'SS), forming an intron lariat. The freed 5' exon remains anchored to loop I of U5 small nuclear RNA (snRNA), and the 5'SS and BPS of the intron form duplexes with conserved U6 and U2 snRNA sequences, respectively. Specific placement of these RNA elements at the catalytic cavity of Prp8 is stabilized by 15 protein components, including Snu114 and the splicing factors Cwc21, Cwc22, Cwc25, and Yju2. These features, representing the conformation of the spliceosome after the first-step reaction, predict structural changes that are needed for the execution of the second-step transesterification reaction. | |||||||||
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_9525.map.gz | 229 MB | EMDB map data format | |
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Header (meta data) | emd-9525-v30.xml emd-9525.xml | 62 KB 62 KB | Display Display | EMDB header |
Images | emd_9525.png | 184.4 KB | ||
Filedesc metadata | emd-9525.cif.gz | 14 KB | ||
Others | emd_9525_additional_1.map.gz emd_9525_additional_2.map.gz | 228.9 MB 228.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-9525 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-9525 | HTTPS FTP |
-Validation report
Summary document | emd_9525_validation.pdf.gz | 639.1 KB | Display | EMDB validaton report |
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Full document | emd_9525_full_validation.pdf.gz | 638.7 KB | Display | |
Data in XML | emd_9525_validation.xml.gz | 7 KB | Display | |
Data in CIF | emd_9525_validation.cif.gz | 8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9525 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9525 | HTTPS FTP |
-Related structure data
Related structure data | 5gmkMC 9526C 9527C 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_9525.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Cryo-EM map of Catalytic Step I spliceosome (C complex) at 3.4 angstrom | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.306 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: Cryo-EM map of Catalytic Step I spliceosome (C...
File | emd_9525_additional_1.map | ||||||||||||
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Annotation | Cryo-EM map of Catalytic Step I spliceosome (C complex) at 3.65 angstrom | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Cryo-EM map of Catalytic Step I spliceosome (C...
File | emd_9525_additional_2.map | ||||||||||||
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Annotation | Cryo-EM map of Catalytic Step I spliceosome (C complex) at 4.6 angstrom | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Spliceosomal C Complex
+Supramolecule #1: Spliceosomal C Complex
+Macromolecule #1: Pre-mRNA-splicing factor 8
+Macromolecule #2: Pre-mRNA-splicing factor SNU114
+Macromolecule #9: Pre-mRNA-splicing factor CWC21
+Macromolecule #10: Pre-mRNA-splicing factor PRP46
+Macromolecule #11: Pre-mRNA-processing protein 45
+Macromolecule #12: Pre-mRNA-splicing factor SLT11
+Macromolecule #13: Pre-mRNA-splicing factor CWC2
+Macromolecule #14: Pre-mRNA-splicing factor CWC15
+Macromolecule #15: Pre-mRNA-splicing factor BUD31
+Macromolecule #16: Pre-mRNA-splicing factor CWC22
+Macromolecule #17: Pre-mRNA-splicing factor CEF1
+Macromolecule #18: Pre-mRNA-splicing factor CLF1
+Macromolecule #19: Protein CWC16
+Macromolecule #20: Pre-mRNA-splicing factor CWC25
+Macromolecule #21: Pre-mRNA-splicing factor ISY1
+Macromolecule #22: Pre-mRNA-splicing factor SYF2
+Macromolecule #23: Pre-mRNA-splicing factor SYF1
+Macromolecule #24: Pre-mRNA-processing factor 17
+Macromolecule #25: Pre-mRNA-processing factor 19
+Macromolecule #26: Pre-mRNA-splicing factor SNT309
+Macromolecule #27: Small nuclear ribonucleoprotein-associated protein B
+Macromolecule #28: Small nuclear ribonucleoprotein E
+Macromolecule #29: Small nuclear ribonucleoprotein F
+Macromolecule #30: Small nuclear ribonucleoprotein G
+Macromolecule #31: Small nuclear ribonucleoprotein Sm D3
+Macromolecule #32: Small nuclear ribonucleoprotein Sm D1
+Macromolecule #33: Small nuclear ribonucleoprotein Sm D2
+Macromolecule #34: U2 small nuclear ribonucleoprotein B''
+Macromolecule #35: U2 small nuclear ribonucleoprotein A'
+Macromolecule #3: U5 snRNA
+Macromolecule #4: U6 snRNA
+Macromolecule #5: U2 snRNA
+Macromolecule #6: Intron_BPS
+Macromolecule #7: 5'-Exon
+Macromolecule #8: 5'-Splicing Site
+Macromolecule #36: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #37: MAGNESIUM ION
+Macromolecule #38: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 8 Details: The CEB buffer (10 mM Tris-HCl, pH 8.0, 75 mM NaCl, 1 mM Mg(OAc)2, 1 mM imidazole, 0.01% NP40, 1 mM TCEP, 0.5 mM EGTA) |
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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) / Digitization - Scanner: OTHER / Average electron dose: 4.7 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 | Resolution.type: BY AUTHOR / Resolution: 3.4 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 161066 |
Initial angle assignment | Type: ANGULAR RECONSTITUTION |
Final angle assignment | Type: ANGULAR RECONSTITUTION |