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- PDB-8h6e: Cryo-EM structure of human exon-defined spliceosome in the late p... -
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Open data
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Basic information
Entry | Database: PDB / ID: 8h6e | ||||||
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Title | Cryo-EM structure of human exon-defined spliceosome in the late pre-B state. | ||||||
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![]() | SPLICING / exon-defined spliceosome / late pre-B state / RNA splicing | ||||||
Function / homology | ![]() Lsm2-8 complex / U6 snRNA 3'-end binding / spliceosomal snRNP complex / ribonucleoprotein complex localization / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / RNA localization / U4atac snRNA binding / R-loop processing ...Lsm2-8 complex / U6 snRNA 3'-end binding / spliceosomal snRNP complex / ribonucleoprotein complex localization / U4atac snRNP / positive regulation of cytotoxic T cell differentiation / maturation of 5S rRNA / RNA localization / U4atac snRNA binding / R-loop processing / mRNA decay by 5' to 3' exoribonuclease / Lsm1-7-Pat1 complex / positive regulation of hippo signaling / regulation of mitotic cell cycle spindle assembly checkpoint / box C/D sno(s)RNA binding / U11/U12 snRNP / U6 snRNP / PH domain binding / U2 snRNP binding / dense fibrillar component / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / histone pre-mRNA 3'end processing complex / cis assembly of pre-catalytic spliceosome / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / box C/D methylation guide snoRNP complex / B-WICH complex / splicing factor binding / U4/U6 snRNP / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / protein methylation / U12-type spliceosomal complex / 7-methylguanosine cap hypermethylation / sno(s)RNA-containing ribonucleoprotein complex / U1 snRNP binding / RNA splicing, via transesterification reactions / methylosome / blastocyst formation / pICln-Sm protein complex / U2-type catalytic step 1 spliceosome / U4 snRNA binding / snRNP binding / small nuclear ribonucleoprotein complex / telomerase RNA binding / telomerase holoenzyme complex / SMN-Sm protein complex / P granule / spliceosomal tri-snRNP complex / U2-type precatalytic spliceosome / U2-type spliceosomal complex / commitment complex / mRNA cis splicing, via spliceosome / U2-type prespliceosome assembly / P-body assembly / U2-type catalytic step 2 spliceosome / SAGA complex / U4 snRNP / U2 snRNP / RNA Polymerase II Transcription Termination / U3 snoRNA binding / U1 snRNP / U2-type prespliceosome / rRNA modification in the nucleus and cytosol / positive regulation of transcription by RNA polymerase III / tRNA processing / precatalytic spliceosome / K63-linked polyubiquitin modification-dependent protein binding / positive regulation of transcription by RNA polymerase I / spliceosomal complex assembly / regulation of RNA splicing / mRNA catabolic process / mRNA Splicing - Minor Pathway / mRNA 3'-splice site recognition / negative regulation of mRNA splicing, via spliceosome / MLL1 complex / nuclear-transcribed mRNA catabolic process / spliceosomal tri-snRNP complex assembly / protein deubiquitination / U5 snRNA binding / U5 snRNP / U2 snRNA binding / U6 snRNA binding / Major pathway of rRNA processing in the nucleolus and cytosol / pre-mRNA intronic binding / RNA processing / regulation of DNA repair / spliceosomal snRNP assembly / ribonucleoprotein complex binding / Cajal body / U1 snRNA binding / U4/U6 x U5 tri-snRNP complex / catalytic step 2 spliceosome / Gene and protein expression by JAK-STAT signaling after Interleukin-12 stimulation / mRNA Splicing - Major Pathway / RNA splicing / positive regulation of protein export from nucleus / helicase activity / maturation of SSU-rRNA Similarity search - Function | ||||||
Biological species | ![]() | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.2 Å | ||||||
![]() | Zhang, W. / Zhan, X. / Zhang, X. / Bai, R. / Lei, J. / Yan, C. / Shi, Y. | ||||||
Funding support | ![]()
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![]() | ![]() Title: Structural insights into human exon-defined spliceosome prior to activation. Authors: Wenyu Zhang / Xiaofeng Zhang / Xiechao Zhan / Rui Bai / Jianlin Lei / Chuangye Yan / Yigong Shi / ![]() Abstract: Spliceosome is often assembled across an exon and undergoes rearrangement to span a neighboring intron. Most states of the intron-defined spliceosome have been structurally characterized. However, ...Spliceosome is often assembled across an exon and undergoes rearrangement to span a neighboring intron. Most states of the intron-defined spliceosome have been structurally characterized. However, the structure of a fully assembled exon-defined spliceosome remains at large. During spliceosome assembly, the pre-catalytic state (B complex) is converted from its precursor (pre-B complex). Here we report atomic structures of the exon-defined human spliceosome in four sequential states: mature pre-B, late pre-B, early B, and mature B. In the previously unknown late pre-B state, U1 snRNP is already released but the remaining proteins are still in the pre-B state; unexpectedly, the RNAs are in the B state, with U6 snRNA forming a duplex with 5'-splice site and U5 snRNA recognizing the 3'-end of the exon. In the early and mature B complexes, the B-specific factors are stepwise recruited and specifically recognize the exon 3'-region. Our study reveals key insights into the assembly of the exon-defined spliceosomes and identifies mechanistic steps of the pre-B-to-B transition. | ||||||
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Structure visualization
Structure viewer | Molecule: ![]() ![]() |
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PDBx/mmCIF format | ![]() | 2.6 MB | Display | ![]() |
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-Validation report
Summary document | ![]() | 949.6 KB | Display | ![]() |
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Full document | ![]() | 984.7 KB | Display | |
Data in XML | ![]() | 223.1 KB | Display | |
Data in CIF | ![]() | 383.1 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 34500MC ![]() 8h6jC ![]() 8h6kC ![]() 8h6lC M: map data used to model this data C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
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Assembly
Deposited unit | ![]()
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Components
+RNA chain , 5 types, 5 molecules A6A5A4A2A
+U6 snRNA-associated Sm-like protein ... , 7 types, 7 molecules 6a6b6c6d6e6f6g
+Protein , 11 types, 13 molecules 5B5C5a4a2a4E4F4G4R4U4X4Y2L
+U5 small nuclear ribonucleoprotein ... , 2 types, 2 molecules 5D5E
+Small nuclear ribonucleoprotein ... , 6 types, 18 molecules 5b4b2b5c4c2c5d4d2d5e4e2e5f4f2f5g4g2g
+U4/U6 small nuclear ribonucleoprotein ... , 3 types, 3 molecules 4B4C4D
+U4/U6.U5 tri-snRNP-associated protein ... , 2 types, 2 molecules 4S4T
+U2 small nuclear ribonucleoprotein ... , 2 types, 2 molecules 2B2C
+Splicing factor 3A subunit ... , 3 types, 3 molecules 2D2E2F
+Splicing factor 3B subunit ... , 6 types, 6 molecules 2G2H2I2J2K2M
+Non-polymers , 4 types, 4 molecules 






+Details
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
Component | Name: exon-defined post pre-B spliceosome / Type: COMPLEX / Entity ID: #2-#21, #1, #22-#47 / Source: NATURAL |
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Molecular weight | Value: 2 MDa / Experimental value: NO |
Source (natural) | Organism: ![]() |
Buffer solution | pH: 8 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
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Electron microscopy imaging
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: ![]() |
Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2500 nm / Nominal defocus min: 1000 nm |
Image recording | Electron dose: 50 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
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Processing
CTF correction | Type: NONE | ||||||||||||||||||||||||
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3D reconstruction | Resolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 140426 / Symmetry type: POINT | ||||||||||||||||||||||||
Refine LS restraints |
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