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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-4255 | |||||||||
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| Title | human Bact spliceosome core structure | |||||||||
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Keywords | spliceosome / human / HELA / BACT / dynamics / SPLICING | |||||||||
| Function / homology | Function and homology informationRES complex / negative regulation of chemokine-mediated signaling pathway / snoRNA splicing / U11/U12 snRNP / regulation of retinoic acid receptor signaling pathway / post-mRNA release spliceosomal complex / regulation of vitamin D receptor signaling pathway / B-WICH complex / nuclear retinoic acid receptor binding / embryonic brain development ...RES complex / negative regulation of chemokine-mediated signaling pathway / snoRNA splicing / U11/U12 snRNP / regulation of retinoic acid receptor signaling pathway / post-mRNA release spliceosomal complex / regulation of vitamin D receptor signaling pathway / B-WICH complex / nuclear retinoic acid receptor binding / embryonic brain development / U12-type spliceosomal complex / mRNA 3'-end processing / RNA splicing, via transesterification reactions / C2H2 zinc finger domain binding / U2-type catalytic step 1 spliceosome / pre-mRNA binding / regulation of mRNA splicing, via spliceosome / positive regulation of mRNA splicing, via spliceosome / blastocyst formation / splicing factor binding / host-mediated activation of viral transcription / U2-type precatalytic spliceosome / positive regulation of vitamin D receptor signaling pathway / mRNA cis splicing, via spliceosome / U2-type spliceosomal complex / nuclear vitamin D receptor binding / U2-type prespliceosome assembly / Notch binding / Transport of Mature mRNA derived from an Intron-Containing Transcript / U2-type catalytic step 2 spliceosome / Regulation of gene expression in late stage (branching morphogenesis) pancreatic bud precursor cells / RUNX3 regulates NOTCH signaling / positive regulation of neurogenesis / NOTCH4 Intracellular Domain Regulates Transcription / RHOBTB1 GTPase cycle / SAGA complex / RNA Polymerase II Transcription Termination / U2 snRNP / NOTCH3 Intracellular Domain Regulates Transcription / U2-type prespliceosome / positive regulation of transcription by RNA polymerase III / WD40-repeat domain binding / protein peptidyl-prolyl isomerization / K63-linked polyubiquitin modification-dependent protein binding / nuclear androgen receptor binding / precatalytic spliceosome / Notch-HLH transcription pathway / Formation of paraxial mesoderm / SMAD binding / positive regulation of transforming growth factor beta receptor signaling pathway / regulation of RNA splicing / spliceosomal complex assembly / mRNA Splicing - Minor Pathway / mRNA 3'-splice site recognition / positive regulation of transcription by RNA polymerase I / spliceosomal tri-snRNP complex assembly / Prp19 complex / U5 snRNA binding / U5 snRNP / intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / protein localization to nucleus / U2 snRNA binding / positive regulation of G1/S transition of mitotic cell cycle / U6 snRNA binding / pre-mRNA intronic binding / RHOBTB2 GTPase cycle / U1 snRNA binding / Cajal body / regulation of DNA repair / retinoic acid receptor signaling pathway / U4/U6 x U5 tri-snRNP complex / cellular response to retinoic acid / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / RNA splicing / DNA damage checkpoint signaling / positive regulation of RNA splicing / positive regulation of protein export from nucleus / nuclear receptor binding / response to cocaine / RNA polymerase II transcription regulatory region sequence-specific DNA binding / stem cell differentiation / peptidylprolyl isomerase / spliceosomal complex / peptidyl-prolyl cis-trans isomerase activity / Downregulation of SMAD2/3:SMAD4 transcriptional activity / positive regulation of neuron projection development / mRNA splicing, via spliceosome / RING-type E3 ubiquitin transferase / negative regulation of protein catabolic process / NOTCH1 Intracellular Domain Regulates Transcription / B-WICH complex positively regulates rRNA expression / positive regulation of protein import into nucleus / Pre-NOTCH Transcription and Translation / Constitutive Signaling by NOTCH1 PEST Domain Mutants / Constitutive Signaling by NOTCH1 HD+PEST Domain Mutants / cellular response to xenobiotic stimulus / nuclear matrix / fibrillar center / mRNA processing Similarity search - Function | |||||||||
| Biological species | Homo sapiens (human) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.4 Å | |||||||||
Authors | Haselbach D / Komarov I / Agafonov D / Hartmuth K / Graf B / Kastner B | |||||||||
Citation | Journal: Cell / Year: 2018Title: Structure and Conformational Dynamics of the Human Spliceosomal B Complex. Authors: David Haselbach / Ilya Komarov / Dmitry E Agafonov / Klaus Hartmuth / Benjamin Graf / Olexandr Dybkov / Henning Urlaub / Berthold Kastner / Reinhard Lührmann / Holger Stark / ![]() Abstract: The spliceosome is a highly dynamic macromolecular complex that precisely excises introns from pre-mRNA. Here we report the cryo-EM 3D structure of the human B spliceosome at 3.4 Å resolution. In ...The spliceosome is a highly dynamic macromolecular complex that precisely excises introns from pre-mRNA. Here we report the cryo-EM 3D structure of the human B spliceosome at 3.4 Å resolution. In the B state, the spliceosome is activated but not catalytically primed, so that it is functionally blocked prior to the first catalytic step of splicing. The spliceosomal core is similar to the yeast B spliceosome; important differences include the presence of the RNA helicase aquarius and peptidyl prolyl isomerases. To examine the overall dynamic behavior of the purified spliceosome, we developed a principal component analysis-based approach. Calculating the energy landscape revealed eight major conformational states, which we refined to higher resolution. Conformational differences of the highly flexible structural components between these eight states reveal how spliceosomal components contribute to the assembly of the spliceosome, allowing it to generate a dynamic interaction network required for its subsequent catalytic activation. | |||||||||
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Structure visualization
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
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Downloads & links
-EMDB archive
| Map data | emd_4255.map.gz | 25.1 MB | EMDB map data format | |
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| Header (meta data) | emd-4255-v30.xml emd-4255.xml | 65 KB 65 KB | Display Display | EMDB header |
| Images | emd_4255.png | 53.1 KB | ||
| Filedesc metadata | emd-4255.cif.gz | 19.7 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-4255 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-4255 | HTTPS FTP |
-Validation report
| Summary document | emd_4255_validation.pdf.gz | 391.8 KB | Display | EMDB validaton report |
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| Full document | emd_4255_full_validation.pdf.gz | 391.3 KB | Display | |
| Data in XML | emd_4255_validation.xml.gz | 7.4 KB | Display | |
| Data in CIF | emd_4255_validation.cif.gz | 8.6 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4255 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4255 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 6ff4MC ![]() 4233C ![]() 4234C ![]() 4235C ![]() 4236C ![]() 4237C ![]() 4238C ![]() 4239C ![]() 4240C ![]() 4247C ![]() 4248C ![]() 4249C ![]() 4250C ![]() 4251C ![]() 4252C ![]() 4253C ![]() 4254C ![]() 6ff7C 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_4255.map.gz / Format: CCP4 / Size: 282.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| 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 : human Bact spliceosome state 1 unmasked
+Supramolecule #1: human Bact spliceosome state 1 unmasked
+Macromolecule #1: RNA-binding motif protein, X-linked 2
+Macromolecule #3: BUD13 homolog
+Macromolecule #6: Splicing factor 3A subunit 2
+Macromolecule #7: Splicing factor 3B subunit 2
+Macromolecule #8: Pre-mRNA-processing-splicing factor 8
+Macromolecule #9: 116 kDa U5 small nuclear ribonucleoprotein component
+Macromolecule #10: SNW domain-containing protein 1
+Macromolecule #11: Pleiotropic regulator 1
+Macromolecule #12: Pre-mRNA-processing factor 17
+Macromolecule #13: Cell division cycle 5-like protein
+Macromolecule #14: Crooked neck-like protein 1
+Macromolecule #15: Pre-mRNA-splicing factor RBM22
+Macromolecule #16: Protein BUD31 homolog
+Macromolecule #17: Spliceosome-associated protein CWC15 homolog
+Macromolecule #18: Serine/arginine repetitive matrix protein 2
+Macromolecule #19: Peptidyl-prolyl cis-trans isomerase-like 1
+Macromolecule #20: Serine/arginine repetitive matrix protein 1
+Macromolecule #22: Peptidyl-prolyl cis-trans isomerase CWC27 homolog
+Macromolecule #23: RING finger protein 113A
+Macromolecule #24: Splicing factor 3B subunit 1
+Macromolecule #25: Splicing factor 3B subunit 3
+Macromolecule #26: Splicing factor 3B subunit 5
+Macromolecule #27: PHD finger-like domain-containing protein 5A
+Macromolecule #28: Splicing factor 3B subunit 6
+Macromolecule #2: U2 snRNA
+Macromolecule #4: U5 snRNA
+Macromolecule #5: U6 snRNA
+Macromolecule #21: pre mRNA
+Macromolecule #29: MAGNESIUM ION
+Macromolecule #30: INOSITOL HEXAKISPHOSPHATE
+Macromolecule #31: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #32: ZINC 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
| Concentration | 0.05 mg/mL | ||||||||||||
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| Buffer | pH: 7.9 Component:
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| Grid | Model: Quantifoil R3.5/1 / Material: COPPER / Mesh: 400 / Support film - #0 - Film type ID: 1 / Support film - #0 - Material: CARBON / Support film - #0 - topology: HOLEY / Support film - #1 - Film type ID: 2 / Support film - #1 - Material: CARBON / Support film - #1 - topology: CONTINUOUS | ||||||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 75 % / Chamber temperature: 277 K / Instrument: LEICA EM GP / Details: blot with blotting sensor. |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Alignment procedure | Coma free - Residual tilt: 14.0 mrad |
| Specialist optics | Spherical aberration corrector: Microscope was modified with a Cs corrector with two hexapoles elements |
| 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 / Number grids imaged: 1 / Number real images: 32000 / Average exposure time: 1.0 sec. / Average electron dose: 40.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Cs: 0.001 mm / Nominal defocus max: 4.5 µm / Nominal defocus min: 0.8 µm / Nominal magnification: 59000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
| Refinement | Protocol: RIGID BODY FIT |
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| Output model | ![]() PDB-6ff4: |
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Keywords
Homo sapiens (human)
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