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Yorodumi- EMDB-6890: Cryo-EM structure of the human activated spliceosome (late Bact) ... -
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Basic information
| Entry | Database: EMDB / ID: EMD-6890 | ||||||||||||
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| Title | Cryo-EM structure of the human activated spliceosome (late Bact) at 6.5 angstrom | ||||||||||||
Map data | Cryo-EM structure of the human activated spliceosome (late Bact) at 6.5 angstrom | ||||||||||||
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Keywords | spliceosome / cryo-EM structure / activated spliceosome / late Bact complex / pre-mRNA splicing / SPLICING | ||||||||||||
| Function / homology | Function and homology informationRES complex / U11/U12 snRNP / post-mRNA release spliceosomal complex / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / generation of catalytic spliceosome for first transesterification step / U7 snRNP / 3'-5' RNA helicase activity / cis assembly of pre-catalytic spliceosome ...RES complex / U11/U12 snRNP / post-mRNA release spliceosomal complex / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / generation of catalytic spliceosome for first transesterification step / U7 snRNP / 3'-5' RNA helicase activity / cis assembly of pre-catalytic spliceosome / histone pre-mRNA 3'end processing complex / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / B-WICH complex / miRNA processing / oocyte development / alternative mRNA splicing, via spliceosome / poly(A) binding / 7-methylguanosine cap hypermethylation / U12-type spliceosomal complex / protein methylation / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U1 snRNP binding / pre-mRNA binding / U2-type catalytic step 1 spliceosome / pICln-Sm protein complex / C2H2 zinc finger domain binding / methylosome / regulation of mRNA splicing, via spliceosome / embryonic brain development / positive regulation of mRNA splicing, via spliceosome / RNA splicing, via transesterification reactions / mRNA 3'-end processing / sno(s)RNA-containing ribonucleoprotein complex / small nuclear ribonucleoprotein complex / SMN-Sm protein complex / spliceosomal tri-snRNP complex / splicing factor binding / blastocyst formation / P granule / snRNP binding / commitment complex / U2-type precatalytic spliceosome / mRNA cis splicing, via spliceosome / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / U2-type spliceosomal complex / Transport of Mature mRNA derived from an Intron-Containing Transcript / telomerase holoenzyme complex / inner cell mass cell proliferation / telomerase RNA binding / U1 snRNP / SAGA complex / U2 snRNP / RNA Polymerase II Transcription Termination / U4 snRNP / U2-type prespliceosome / positive regulation of transcription by RNA polymerase III / protein peptidyl-prolyl isomerization / K63-linked polyubiquitin modification-dependent protein binding / mRNA stabilization / cyclosporin A binding / ubiquitin-ubiquitin ligase activity / precatalytic spliceosome / lipid biosynthetic process / pattern recognition receptor activity / WD40-repeat domain binding / mRNA 3'-splice site recognition / blastocyst development / regulation of RNA splicing / mRNA Splicing - Minor Pathway / positive regulation of transcription by RNA polymerase I / Dengue Virus-Host Interactions / spliceosomal complex assembly / negative regulation of mRNA splicing, via spliceosome / spliceosomal tri-snRNP complex assembly / Prp19 complex / U5 snRNP / U5 snRNA binding / pre-mRNA intronic binding / U2 snRNA binding / U6 snRNA binding / protein K63-linked ubiquitination / spliceosomal snRNP assembly / protein localization to nucleus / Cajal body / U1 snRNA binding / regulation of DNA repair / RNA processing / positive regulation of G1/S transition of mitotic cell cycle / U4/U6 x U5 tri-snRNP complex / transcription regulator inhibitor activity / positive regulation of viral genome replication / ovarian follicle development / spindle assembly / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / DNA damage checkpoint signaling / lipid droplet / transcription-coupled nucleotide-excision repair Similarity search - Function | ||||||||||||
| Biological species | Homo sapiens (human) / unidentified adenovirus | ||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 6.5 Å | ||||||||||||
Authors | Zhang X / Yan C | ||||||||||||
| Funding support | China, 3 items
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Citation | Journal: Cell Res / Year: 2018Title: Structure of the human activated spliceosome in three conformational states. Authors: Xiaofeng Zhang / Chuangye Yan / Xiechao Zhan / Lijia Li / Jianlin Lei / Yigong Shi / ![]() Abstract: During each cycle of pre-mRNA splicing, the pre-catalytic spliceosome (B complex) is converted into the activated spliceosome (B complex), which has a well-formed active site but cannot proceed to ...During each cycle of pre-mRNA splicing, the pre-catalytic spliceosome (B complex) is converted into the activated spliceosome (B complex), which has a well-formed active site but cannot proceed to the branching reaction. Here, we present the cryo-EM structure of the human B complex in three distinct conformational states. The EM map allows atomic modeling of nearly all protein components of the U2 small nuclear ribonucleoprotein (snRNP), including three of the SF3a complex and seven of the SF3b complex. The structure of the human B complex contains 52 proteins, U2, U5, and U6 small nuclear RNA (snRNA), and a pre-mRNA. Three distinct conformations have been captured, representing the early, mature, and late states of the human B complex. These complexes differ in the orientation of the Switch loop of Prp8, the splicing factors RNF113A and NY-CO-10, and most components of the NineTeen complex (NTC) and the NTC-related complex. Analysis of these three complexes and comparison with the B and C complexes reveal an ordered flux of components in the B-to-B and the B-to-B transitions, which ultimately prime the active site for the branching reaction. | ||||||||||||
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Structure visualization
| Movie |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_6890.map.gz | 228.2 MB | EMDB map data format | |
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| Header (meta data) | emd-6890-v30.xml emd-6890.xml | 78.8 KB 78.8 KB | Display Display | EMDB header |
| Images | emd_6890.png | 171.8 KB | ||
| Filedesc metadata | emd-6890.cif.gz | 24.4 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-6890 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-6890 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 5z57MC ![]() 6889C ![]() 6891C ![]() 5z56C ![]() 5z58C 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_6890.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | Cryo-EM structure of the human activated spliceosome (late Bact) at 6.5 angstrom | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.338 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 : late Bact splieosome
+Supramolecule #1: late Bact splieosome
+Macromolecule #1: Pre-mRNA-processing-splicing factor 8
+Macromolecule #3: 116 kDa U5 small nuclear ribonucleoprotein component
+Macromolecule #4: U5 small nuclear ribonucleoprotein 200 kDa helicase
+Macromolecule #5: U5 small nuclear ribonucleoprotein 40 kDa protein
+Macromolecule #6: Small nuclear ribonucleoprotein Sm D3
+Macromolecule #7: Small nuclear ribonucleoprotein-associated proteins B and B'
+Macromolecule #8: Small nuclear ribonucleoprotein Sm D1
+Macromolecule #9: Small nuclear ribonucleoprotein Sm D2
+Macromolecule #10: Small nuclear ribonucleoprotein F
+Macromolecule #11: Small nuclear ribonucleoprotein E
+Macromolecule #12: Small nuclear ribonucleoprotein G
+Macromolecule #16: U2 small nuclear ribonucleoprotein A'
+Macromolecule #17: U2 small nuclear ribonucleoprotein B''
+Macromolecule #18: Splicing factor 3A subunit 3
+Macromolecule #19: Splicing factor 3A subunit 1
+Macromolecule #20: Splicing factor 3A subunit 2
+Macromolecule #21: Splicing factor 3B subunit 1
+Macromolecule #22: Splicing factor 3B subunit 2
+Macromolecule #23: Splicing factor 3B subunit 3
+Macromolecule #24: Splicing factor 3B subunit 4
+Macromolecule #25: Splicing factor 3B subunit 6
+Macromolecule #26: PHD finger-like domain-containing protein 5A
+Macromolecule #27: Splicing factor 3B subunit 5
+Macromolecule #28: Crooked neck-like protein 1
+Macromolecule #29: Cell division cycle 5-like protein
+Macromolecule #30: Pre-mRNA-processing factor 19
+Macromolecule #31: Pre-mRNA-splicing factor SPF27
+Macromolecule #32: Pre-mRNA-splicing factor SYF1
+Macromolecule #33: Intron-binding protein aquarius
+Macromolecule #34: Protein BUD31 homolog
+Macromolecule #35: Pre-mRNA-splicing factor RBM22
+Macromolecule #36: Spliceosome-associated protein CWC15 homolog
+Macromolecule #37: Skip
+Macromolecule #38: Peptidyl-prolyl cis-trans isomerase-like 1
+Macromolecule #39: Pleiotropic regulator 1
+Macromolecule #40: Serine/arginine repetitive matrix protein 2
+Macromolecule #41: Pre-mRNA-splicing factor CWC22 homolog
+Macromolecule #42: Pre-mRNA-processing factor 17
+Macromolecule #43: Smad nuclear-interacting protein 1
+Macromolecule #44: RNA-binding motif protein, X-linked 2
+Macromolecule #45: BUD13 homolog
+Macromolecule #46: Pre-mRNA-splicing factor ATP-dependent RNA helicase DHX16
+Macromolecule #47: Peptidyl-prolyl cis-trans isomerase E
+Macromolecule #2: U5 snRNA
+Macromolecule #13: U6 snRNA
+Macromolecule #14: pre-mRNA
+Macromolecule #15: U2 snRNA
+Macromolecule #48: INOSITOL HEXAKISPHOSPHATE
+Macromolecule #49: ALANINE
+Macromolecule #50: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #51: MAGNESIUM ION
+Macromolecule #52: 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
| Buffer | pH: 7.9 / Details: 20 mM HEPES-KOH, pH 7.9, 150 mM NaCl, 1.5 mM MgCl2 |
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| Vitrification | Cryogen name: ETHANE |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Specialist optics | Energy filter - Name: GIF Quantum LS |
| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Average electron dose: 48.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm |
| 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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About Yorodumi


Keywords
Homo sapiens (human)
unidentified adenovirus
Authors
China, 3 items
Citation
UCSF Chimera









































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