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- EMDB-6889: cryo-EM structure of a human activated spliceosome (mature Bact) ... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-6889 | ||||||||||||
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Title | cryo-EM structure of a human activated spliceosome (mature Bact) at 5.1 angstrom. | ||||||||||||
![]() | Cryo-EM structure of the human activated spliceosome (mature Bact) at 5.1 angstrom resolution | ||||||||||||
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![]() | human activated spliceosome / mature Bact / pre-mRNA splicing / SPLICING | ||||||||||||
Function / homology | ![]() RES complex / negative regulation of chemokine-mediated signaling pathway / snoRNA splicing / U11/U12 snRNP / post-mRNA release spliceosomal complex / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / 3'-5' RNA helicase activity ...RES complex / negative regulation of chemokine-mediated signaling pathway / snoRNA splicing / U11/U12 snRNP / post-mRNA release spliceosomal complex / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / 3'-5' RNA helicase activity / generation of catalytic spliceosome for first transesterification step / histone pre-mRNA 3'end processing complex / alternative mRNA splicing, via spliceosome / cis assembly of pre-catalytic spliceosome / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / B-WICH complex / miRNA processing / splicing factor binding / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / embryonic brain development / protein methylation / U12-type spliceosomal complex / poly(A) binding / 7-methylguanosine cap hypermethylation / U1 snRNP binding / RNA splicing, via transesterification reactions / mRNA 3'-end processing / sno(s)RNA-containing ribonucleoprotein complex / methylosome / blastocyst formation / pICln-Sm protein complex / regulation of mRNA splicing, via spliceosome / oocyte development / C2H2 zinc finger domain binding / U2-type catalytic step 1 spliceosome / positive regulation of mRNA splicing, via spliceosome / pre-mRNA binding / snRNP binding / small nuclear ribonucleoprotein complex / telomerase holoenzyme complex / SMN-Sm protein complex / P granule / telomerase RNA binding / spliceosomal tri-snRNP complex / U2-type precatalytic spliceosome / U2-type spliceosomal complex / commitment complex / mRNA cis splicing, via spliceosome / U2-type prespliceosome assembly / Transport of Mature mRNA derived from an Intron-Containing Transcript / U2-type catalytic step 2 spliceosome / SAGA complex / U4 snRNP / U2 snRNP / RNA Polymerase II Transcription Termination / U1 snRNP / U2-type prespliceosome / WD40-repeat domain binding / protein peptidyl-prolyl isomerization / positive regulation of transcription by RNA polymerase III / inner cell mass cell proliferation / ubiquitin-ubiquitin ligase activity / K63-linked polyubiquitin modification-dependent protein binding / precatalytic spliceosome / lipid biosynthetic process / cyclosporin A binding / pattern recognition receptor activity / positive regulation of transcription by RNA polymerase I / spliceosomal complex assembly / regulation of RNA splicing / mRNA Splicing - Minor Pathway / mRNA 3'-splice site recognition / spliceosomal tri-snRNP complex assembly / blastocyst development / Prp19 complex / protein K63-linked ubiquitination / U5 snRNA binding / protein localization to nucleus / U5 snRNP / positive regulation of G1/S transition of mitotic cell cycle / U2 snRNA binding / U6 snRNA binding / pre-mRNA intronic binding / positive regulation of viral genome replication / spliceosomal snRNP assembly / RNA processing / regulation of DNA repair / Cajal body / transcription regulator inhibitor activity / proteasomal protein catabolic process / transcription-coupled nucleotide-excision repair / U1 snRNA binding / spindle assembly / ovarian follicle development / U4/U6 x U5 tri-snRNP complex / negative regulation of canonical NF-kappaB signal transduction / lipid droplet / catalytic step 2 spliceosome / antiviral innate immune response Similarity search - Function | ||||||||||||
Biological species | ![]() ![]() | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 5.1 Å | ||||||||||||
![]() | Zhang X / Yan C | ||||||||||||
Funding support | ![]()
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![]() | ![]() Title: 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. | ||||||||||||
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 | ![]() | 228.5 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 83 KB 83 KB | Display Display | ![]() |
Images | ![]() | 171.8 KB | ||
Filedesc metadata | ![]() | 25 KB | ||
Others | ![]() | 59.6 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 5z56MC ![]() 6890C ![]() 6891C ![]() 5z57C ![]() 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 | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Cryo-EM structure of the human activated spliceosome (mature Bact) at 5.1 angstrom resolution | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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
-Additional map: local map for SF3b region of the human...
File | emd_6889_additional.map | ||||||||||||
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Annotation | local map for SF3b region of the human activated spliceosome at 4.2 angstrom resolution | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
+Entire : human activated spliceosome
+Supramolecule #1: human activated spliceosome
+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: RING finger protein 113A
+Macromolecule #35: Protein BUD31 homolog
+Macromolecule #36: Pre-mRNA-splicing factor RBM22
+Macromolecule #37: Spliceosome-associated protein CWC15 homolog
+Macromolecule #38: Skip
+Macromolecule #39: Peptidyl-prolyl cis-trans isomerase-like 1
+Macromolecule #40: Pleiotropic regulator 1
+Macromolecule #41: Serine/arginine repetitive matrix protein 2
+Macromolecule #42: Pre-mRNA-splicing factor CWC22 homolog
+Macromolecule #43: Pre-mRNA-processing factor 17
+Macromolecule #44: Smad nuclear-interacting protein 1
+Macromolecule #45: RNA-binding motif protein, X-linked 2
+Macromolecule #46: BUD13 homolog
+Macromolecule #47: Peptidyl-prolyl cis-trans isomerase CWC27 homolog
+Macromolecule #48: Putative pre-mRNA-splicing factor ATP-dependent RNA helicase DHX16
+Macromolecule #49: Peptidyl-prolyl cis-trans isomerase E
+Macromolecule #2: U5 snRNA
+Macromolecule #13: U6 snRNA
+Macromolecule #14: pre-mRNA
+Macromolecule #15: U2 snRNA
+Macromolecule #50: INOSITOL HEXAKISPHOSPHATE
+Macromolecule #51: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #52: MAGNESIUM ION
+Macromolecule #53: 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: 7.9 / Details: 20 mM HEPES-KOH, 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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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: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |