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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 / regulation of vitamin D receptor signaling pathway / regulation of retinoic acid receptor signaling pathway / B-WICH complex / C2H2 zinc finger domain binding / U11/U12 snRNP / U4atac/U6atac x U5 tri-snRNP complex / embryonic brain development / U12-type spliceosomal complex ...RES complex / negative regulation of chemokine-mediated signaling pathway / regulation of vitamin D receptor signaling pathway / regulation of retinoic acid receptor signaling pathway / B-WICH complex / C2H2 zinc finger domain binding / U11/U12 snRNP / U4atac/U6atac x U5 tri-snRNP complex / embryonic brain development / U12-type spliceosomal complex / nuclear retinoic acid receptor binding / regulation of mRNA splicing, via spliceosome / post-spliceosomal complex / positive regulation of mRNA splicing, via spliceosome / RNA splicing, via transesterification reactions / mRNA 3'-end processing / U12-type catalytic step 2 spliceosome / post-mRNA release spliceosomal complex / splicing factor binding / U12-type precatalytic spliceosome / positive regulation of vitamin D receptor signaling pathway / RNA Polymerase II Transcription Termination / host-mediated activation of viral transcription / U2-type precatalytic spliceosome / Regulation of gene expression in late stage (branching morphogenesis) pancreatic bud precursor cells / RUNX3 regulates NOTCH signaling / Notch binding / U2-type spliceosomal complex / nuclear vitamin D receptor binding / U2-type prespliceosome assembly / Transport of Mature mRNA derived from an Intron-Containing Transcript / NOTCH4 Intracellular Domain Regulates Transcription / pre-mRNA binding / U2-type catalytic step 2 spliceosome / positive regulation of neurogenesis / SAGA complex / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U2 snRNP / NOTCH3 Intracellular Domain Regulates Transcription / U2-type catalytic step 1 spliceosome / mRNA stabilization / U2-type prespliceosome / mRNA cis splicing, via spliceosome / RHOBTB1 GTPase cycle / protein peptidyl-prolyl isomerization / nuclear androgen receptor binding / K63-linked polyubiquitin modification-dependent protein binding / Notch-HLH transcription pathway / Formation of paraxial mesoderm / precatalytic spliceosome / positive regulation of transforming growth factor beta receptor signaling pathway / regulation of RNA splicing / WD40-repeat domain binding / mRNA 3'-splice site recognition / SMAD binding / positive regulation of transcription by RNA polymerase III / mRNA Splicing - Minor Pathway / spliceosomal complex assembly / positive regulation of transcription by RNA polymerase I / intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / Prp19 complex / negative regulation of mRNA splicing, via spliceosome / U2 snRNA binding / U5 snRNA binding / spliceosomal tri-snRNP complex assembly / U5 snRNP / U6 snRNA binding / U4/U6 x U5 tri-snRNP complex / Cajal body / retinoic acid receptor signaling pathway / RHOBTB2 GTPase cycle / cellular response to retinoic acid / CHD1 and CHD2 subfamily / DNA damage checkpoint signaling / spliceosomal snRNP assembly / stem cell differentiation / positive regulation of protein export from nucleus / catalytic step 2 spliceosome / RNA splicing / spliceosomal complex / mRNA Splicing - Major Pathway / regulation of DNA repair / positive regulation of RNA splicing / nuclear receptor binding / cellular response to xenobiotic stimulus / response to cocaine / RNA polymerase II transcription regulatory region sequence-specific DNA binding / cellular response to tumor necrosis factor / positive regulation of neuron projection development / peptidylprolyl isomerase / peptidyl-prolyl cis-trans isomerase activity / negative regulation of protein catabolic process / positive regulation of protein import into nucleus / mRNA splicing, via spliceosome / Downregulation of SMAD2/3:SMAD4 transcriptional activity / RING-type E3 ubiquitin transferase / fibrillar center / mRNA processing / B-WICH complex positively regulates rRNA expression / NOTCH1 Intracellular Domain Regulates Transcription 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 | https://data.pdbj.org/pub/emdb/structures/EMD-4255 ftp://data.pdbj.org/pub/emdb/structures/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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