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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-11695 | |||||||||
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Title | Human pre-Bact-1 spliceosome | |||||||||
![]() | Unmasked/unsharpened map of the pre-Bact-1 complex. | |||||||||
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![]() | Complex / spliceosome / catalytic activation / splicing | |||||||||
Function / homology | ![]() positive regulation of RNA binding / snRNA export from nucleus / DNA topoisomerase binding / nuclear cap binding complex / histone mRNA metabolic process / RNA cap binding complex / microfibril / mRNA metabolic process / Lsm2-8 complex / U6 snRNA 3'-end binding ...positive regulation of RNA binding / snRNA export from nucleus / DNA topoisomerase binding / nuclear cap binding complex / histone mRNA metabolic process / RNA cap binding complex / microfibril / mRNA metabolic process / Lsm2-8 complex / U6 snRNA 3'-end binding / positive regulation of mRNA 3'-end processing / positive regulation of RNA export from nucleus / protein kinase B binding / mRNA decay by 5' to 3' exoribonuclease / cap-dependent translational initiation / Lsm1-7-Pat1 complex / U11/U12 snRNP / Processing of Intronless Pre-mRNAs / U6 snRNP / PH domain binding / regulation of retinoic acid receptor signaling pathway / snRNA binding / U2 snRNP binding / RNA cap binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / histone pre-mRNA 3'end processing complex / alternative mRNA splicing, via spliceosome / cis assembly of pre-catalytic spliceosome / regulation of vitamin D receptor signaling pathway / primary miRNA processing / miRNA-mediated post-transcriptional gene silencing / mRNA splice site recognition / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / regulatory ncRNA-mediated post-transcriptional gene silencing / regulation of mRNA processing / B-WICH complex / miRNA processing / splicing factor binding / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / nuclear retinoic acid receptor binding / RNA 7-methylguanosine cap binding / protein methylation / U12-type spliceosomal complex / Transport of the SLBP independent Mature mRNA / Transport of the SLBP Dependant Mature mRNA / 7-methylguanosine cap hypermethylation / transcription elongation factor activity / U1 snRNP binding / RNA splicing, via transesterification reactions / mRNA 3'-end processing / sno(s)RNA-containing ribonucleoprotein complex / methylosome / Transport of Mature mRNA Derived from an Intronless Transcript / blastocyst formation / pICln-Sm protein complex / regulation of mRNA splicing, via spliceosome / U2-type catalytic step 1 spliceosome / positive regulation of mRNA splicing, via spliceosome / pre-mRNA binding / snRNP binding / small nuclear ribonucleoprotein complex / mRNA 3'-end processing / telomerase holoenzyme complex / SMN-Sm protein complex / P granule / telomerase RNA binding / spliceosomal tri-snRNP complex / positive regulation by host of viral transcription / U2-type precatalytic spliceosome / U2-type spliceosomal complex / positive regulation of vitamin D receptor signaling pathway / commitment complex / mRNA cis splicing, via spliceosome / U2-type prespliceosome assembly / RNA catabolic process / Transport of Mature mRNA derived from an Intron-Containing Transcript / nuclear vitamin D receptor binding / RNA polymerase binding / Notch binding / P-body assembly / Regulation of gene expression in late stage (branching morphogenesis) pancreatic bud precursor cells / regulation of translational initiation / U2-type catalytic step 2 spliceosome / RUNX3 regulates NOTCH signaling / NOTCH4 Intracellular Domain Regulates Transcription / SAGA complex / U4 snRNP / RHOBTB1 GTPase cycle / Abortive elongation of HIV-1 transcript in the absence of Tat / U2 snRNP / RNA Polymerase II Transcription Termination / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / positive regulation of protein targeting to mitochondrion / U1 snRNP / NOTCH3 Intracellular Domain Regulates Transcription / FGFR2 alternative splicing / ubiquitin-like protein conjugating enzyme binding Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 7.8 Å | |||||||||
![]() | Townsend C / Kastner B | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Mechanism of protein-guided folding of the active site U2/U6 RNA during spliceosome activation. Authors: Cole Townsend / Majety N Leelaram / Dmitry E Agafonov / Olexandr Dybkov / Cindy L Will / Karl Bertram / Henning Urlaub / Berthold Kastner / Holger Stark / Reinhard Lührmann / ![]() Abstract: Spliceosome activation involves extensive protein and RNA rearrangements that lead to formation of a catalytically active U2/U6 RNA structure. At present, little is known about the assembly pathway ...Spliceosome activation involves extensive protein and RNA rearrangements that lead to formation of a catalytically active U2/U6 RNA structure. At present, little is known about the assembly pathway of the latter and the mechanism whereby proteins aid its proper folding. Here, we report the cryo-electron microscopy structures of two human, activated spliceosome precursors (that is, pre-B complexes) at core resolutions of 3.9 and 4.2 angstroms. These structures elucidate the order of the numerous protein exchanges that occur during activation, the mutually exclusive interactions that ensure the correct order of ribonucleoprotein rearrangements needed to form the U2/U6 catalytic RNA, and the stepwise folding pathway of the latter. Structural comparisons with mature B complexes reveal the molecular mechanism whereby a conformational change in the scaffold protein PRP8 facilitates final three-dimensional folding of the U2/U6 catalytic RNA. | |||||||||
History |
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Structure visualization
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 170.7 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 83.8 KB 83.8 KB | Display Display | ![]() |
Images | ![]() | 41.1 KB | ||
Filedesc metadata | ![]() | 22 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7abgMC ![]() 7aavC ![]() 7abfC ![]() 7abhC ![]() 7abiC C: citing same article ( M: atomic model generated by this map |
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Similar structure data | |
EM raw data | ![]() Data #1: Motion-corrected micrographs (without dose-weighting) of human pre-Bact spliceosome [micrographs - single frame] Data #2: Motion-corrected micrographs (with dose-weighting) of human pre-Bact spliceosome [micrographs - single frame]) |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Unmasked/unsharpened map of the pre-Bact-1 complex. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.16 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
+Entire : pre-Bact-1 spliceosomal complex
+Supramolecule #1: pre-Bact-1 spliceosomal complex
+Supramolecule #2: pre-Bact-1 spliceosomal complex
+Macromolecule #1: Nuclear cap-binding protein subunit 1
+Macromolecule #2: U6 snRNA-associated Sm-like protein LSm7
+Macromolecule #3: Splicing factor 3B subunit 6
+Macromolecule #4: Splicing factor 3A subunit 2
+Macromolecule #5: U5 small nuclear ribonucleoprotein 40 kDa protein
+Macromolecule #6: U2 small nuclear ribonucleoprotein A'
+Macromolecule #7: U2 small nuclear ribonucleoprotein B''
+Macromolecule #8: U5 small nuclear ribonucleoprotein 200 kDa helicase
+Macromolecule #9: Protein BUD31 homolog
+Macromolecule #10: Nuclear cap-binding protein subunit 2
+Macromolecule #12: Cell division cycle 5-like protein
+Macromolecule #13: Spliceosome-associated protein CWC15 homolog
+Macromolecule #14: U6 snRNA-associated Sm-like protein LSm2
+Macromolecule #15: U6 snRNA-associated Sm-like protein LSm3
+Macromolecule #16: U6 snRNA-associated Sm-like protein LSm4
+Macromolecule #17: U6 snRNA-associated Sm-like protein LSm5
+Macromolecule #18: U6 snRNA-associated Sm-like protein LSm6
+Macromolecule #20: U6 snRNA-associated Sm-like protein LSm8
+Macromolecule #21: Microfibrillar-associated protein 1
+Macromolecule #22: PHD finger-like domain-containing protein 5A
+Macromolecule #23: Pleiotropic regulator 1
+Macromolecule #25: Pre-mRNA-processing-splicing factor 8
+Macromolecule #26: Pre-mRNA-splicing factor 38A
+Macromolecule #27: Pre-mRNA-splicing factor RBM22
+Macromolecule #28: Splicing factor 3A subunit 1
+Macromolecule #29: Splicing factor 3A subunit 3
+Macromolecule #30: Transcription elongation regulator 1
+Macromolecule #31: Splicing factor 3B subunit 1
+Macromolecule #32: Splicing factor 3B subunit 2
+Macromolecule #33: Splicing factor 3B subunit 3
+Macromolecule #34: Splicing factor 3B subunit 4
+Macromolecule #35: Splicing factor 3B subunit 5
+Macromolecule #36: SNW domain-containing protein 1
+Macromolecule #37: Smad nuclear-interacting protein 1
+Macromolecule #38: Zinc finger matrin-type protein 2
+Macromolecule #39: 116 kDa U5 small nuclear ribonucleoprotein component
+Macromolecule #40: Serine/arginine repetitive matrix protein 1
+Macromolecule #42: Serine/arginine-rich splicing factor 1
+Macromolecule #43: Small nuclear ribonucleoprotein Sm D2
+Macromolecule #44: Small nuclear ribonucleoprotein F
+Macromolecule #45: Small nuclear ribonucleoprotein-associated proteins B and B'
+Macromolecule #46: Small nuclear ribonucleoprotein Sm D3
+Macromolecule #47: Small nuclear ribonucleoprotein G
+Macromolecule #48: Small nuclear ribonucleoprotein E
+Macromolecule #49: Small nuclear ribonucleoprotein Sm D1
+Macromolecule #50: Ubiquitin-like protein 5
+Macromolecule #51: WW domain-binding protein 11
+Macromolecule #11: U5 snRNA
+Macromolecule #19: U2 snRNA
+Macromolecule #24: MINX M3 pre-mRNA
+Macromolecule #41: U6 snRNA
+Macromolecule #52: INOSITOL HEXAKISPHOSPHATE
+Macromolecule #53: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #54: MAGNESIUM ION
+Macromolecule #55: 7-METHYL-GUANOSINE-5'-TRIPHOSPHATE-5'-GUANOSINE
-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: 7.9 |
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Grid | Model: Quantifoil R3.5/1 / Material: COPPER / Support film - Material: CARBON / Support film - topology: CONTINUOUS |
Vitrification | Cryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Detector mode: INTEGRATING / Digitization - Dimensions - Width: 4096 pixel / Digitization - Dimensions - Height: 4096 pixel / Average exposure time: 1.0 sec. / Average electron dose: 2.25 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Startup model | Type of model: OTHER / Details: cryoSPARC ab initio reconstruction |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 7.8 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.0) / Number images used: 84539 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: RIGID BODY FIT |
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Output model | ![]() PDB-7abg: |