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- EMDB-9525: Cryo-EM structure of the Catalytic Step I spliceosome (C complex)... -
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Open data
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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 | |||||||||
![]() | Cryo-EM map of Catalytic Step I spliceosome (C complex) at 3.4 angstrom | |||||||||
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![]() | RNA splicing / Spliceosome / Catalytic Step I / Intron lariat / RNA BINDING PROTEIN-RNA complex | |||||||||
Function / homology | ![]() 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 / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U4/U6 snRNP ...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 / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U4/U6 snRNP / 7-methylguanosine cap hypermethylation / pICln-Sm protein complex / U2-type catalytic step 1 spliceosome / pre-mRNA binding / snRNP binding / small nuclear ribonucleoprotein complex / SMN-Sm protein complex / spliceosomal tri-snRNP complex / U2-type spliceosomal complex / commitment complex / mRNA cis splicing, via spliceosome / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / U4 snRNP / U2 snRNP / U1 snRNP / poly(U) RNA binding / U2-type prespliceosome / precatalytic spliceosome / generation of catalytic spliceosome for second transesterification step / Formation of TC-NER Pre-Incision Complex / spliceosomal complex assembly / mRNA 5'-splice site recognition / Gap-filling DNA repair synthesis and ligation in TC-NER / mRNA 3'-splice site recognition / DNA replication origin binding / Dual incision in TC-NER / spliceosomal tri-snRNP complex assembly / Prp19 complex / protein K63-linked ubiquitination / U5 snRNA binding / DNA replication initiation / U5 snRNP / U2 snRNA binding / U6 snRNA binding / pre-mRNA intronic binding / spliceosomal snRNP assembly / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / positive regulation of cell cycle / catalytic step 2 spliceosome / nuclear periphery / RNA splicing / positive regulation of RNA splicing / spliceosomal complex / RING-type E3 ubiquitin transferase / mRNA splicing, via spliceosome / metallopeptidase activity / ubiquitin-protein transferase activity / ubiquitin protein ligase activity / DNA repair / GTPase activity / mRNA binding / chromatin binding / GTP binding / chromatin / mitochondrion / DNA binding / RNA binding / zinc ion binding / metal ion binding / identical protein binding / nucleus / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | ![]() ![]() ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.4 Å | |||||||||
![]() | Wan R / Yan C | |||||||||
![]() | ![]() 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 |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 229 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 62 KB 62 KB | Display Display | ![]() |
Images | ![]() | 184.4 KB | ||
Filedesc metadata | ![]() | 14 KB | ||
Others | ![]() ![]() | 228.9 MB 228.8 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 5gmkMC ![]() 9526C ![]() 9527C 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 | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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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 |
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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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![]() | single particle reconstruction |
Aggregation state | particle |
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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 |
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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) / Digitization - Scanner: OTHER / Average electron dose: 4.7 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
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: 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 |