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Yorodumi- PDB-5z58: Cryo-EM structure of a human activated spliceosome (early Bact) a... -
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Basic information
| Entry | Database: PDB / ID: 5z58 | ||||||||||||
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| Title | Cryo-EM structure of a human activated spliceosome (early Bact) at 4.9 angstrom. | ||||||||||||
Components |
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Keywords | SPLICING / spliceosome / cryo-EM structure / activated spliceosome / early Bact complex / pre-mRNA splicing | ||||||||||||
| Function / homology | Function and homology informationRES complex / negative regulation of chemokine-mediated signaling pathway / snoRNA splicing / regulation of vitamin D receptor signaling pathway / U11/U12 snRNP / regulation of retinoic acid receptor signaling pathway / post-mRNA release spliceosomal complex / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding ...RES complex / negative regulation of chemokine-mediated signaling pathway / snoRNA splicing / regulation of vitamin D receptor signaling pathway / U11/U12 snRNP / regulation of retinoic acid receptor signaling pathway / post-mRNA release spliceosomal complex / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / 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 / 7-methylguanosine cap hypermethylation / U12-type spliceosomal complex / protein methylation / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / nuclear retinoic acid receptor binding / U1 snRNP binding / U2-type catalytic step 1 spliceosome / pICln-Sm protein complex / methylosome / regulation of mRNA splicing, via spliceosome / positive regulation of mRNA splicing, via spliceosome / RNA splicing, via transesterification reactions / 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 / positive regulation of vitamin D receptor signaling pathway / snRNP binding / commitment complex / host-mediated activation of viral transcription / U2-type precatalytic spliceosome / mRNA cis splicing, via spliceosome / Regulation of gene expression in late stage (branching morphogenesis) pancreatic bud precursor cells / RUNX3 regulates NOTCH signaling / Notch binding / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / U2-type spliceosomal complex / nuclear vitamin D receptor binding / NOTCH4 Intracellular Domain Regulates Transcription / telomerase holoenzyme complex / telomerase RNA binding / U1 snRNP / SAGA complex / U2 snRNP / RNA Polymerase II Transcription Termination / NOTCH3 Intracellular Domain Regulates Transcription / U4 snRNP / U2-type prespliceosome / positive regulation of transcription by RNA polymerase III / K63-linked polyubiquitin modification-dependent protein binding / mRNA stabilization / positive regulation of neurogenesis / nuclear androgen receptor binding / precatalytic spliceosome / Notch-HLH transcription pathway / Formation of paraxial mesoderm / pattern recognition receptor activity / WD40-repeat domain binding / positive regulation of transforming growth factor beta receptor signaling pathway / mRNA 3'-splice site recognition / SMAD binding / 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 / intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / pre-mRNA intronic binding / U2 snRNA binding / U6 snRNA binding / spliceosomal snRNP assembly / protein localization to nucleus / Cajal body / U1 snRNA binding / regulation of DNA repair / RNA processing / retinoic acid receptor signaling pathway / positive regulation of G1/S transition of mitotic cell cycle / U4/U6 x U5 tri-snRNP complex / transcription regulator inhibitor activity / cellular response to retinoic acid / catalytic step 2 spliceosome / mRNA Splicing - Major Pathway / DNA damage checkpoint signaling Similarity search - Function | ||||||||||||
| Biological species | Homo sapiens (human) unidentified adenovirus | ||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 4.9 Å | ||||||||||||
Authors | Zhang, X. / Yan, C. / Zhan, X. / Li, L. / Lei, J. / Shi, Y. | ||||||||||||
| Funding support | China, 3items
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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
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| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 5z58.cif.gz | 2.7 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb5z58.ent.gz | 1.9 MB | Display | PDB format |
| PDBx/mmJSON format | 5z58.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/z5/5z58 ftp://data.pdbj.org/pub/pdb/validation_reports/z5/5z58 | HTTPS FTP |
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-Related structure data
| Related structure data | ![]() 6891MC ![]() 6889C ![]() 6890C ![]() 5z56C ![]() 5z57C C: citing same article ( M: map data used to model this data |
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| Similar structure data |
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Links
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Assembly
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Components
-Protein , 14 types, 15 molecules ACbi6JLMPRTXYZz
| #1: Protein | Mass: 273974.250 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q6P2Q9 | ||||||||||||||||||||||
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| #3: Protein | Mass: 109560.625 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q15029 | ||||||||||||||||||||||
| #7: Protein | Mass: 23686.004 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P14678#26: Protein | | Mass: 12427.524 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q7RTV0#28: Protein | | Mass: 100610.008 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9BZJ0#29: Protein | | Mass: 92406.883 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q99459#30: Protein | | Mass: 38847.199 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O15541#31: Protein | | Mass: 26674.447 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9P013#32: Protein | | Mass: 62063.160 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q13573*PLUS#33: Protein | | Mass: 57280.758 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O43660#35: Protein | | Mass: 45880.738 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q8TAD8#36: Protein | | Mass: 37425.984 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9Y388#37: Protein | | Mass: 70669.211 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9BRD0#38: Protein | | Mass: 53941.227 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q6UX04, peptidylprolyl isomerase |
-RNA chain , 4 types, 4 molecules BFGH
| #2: RNA chain | Mass: 37254.855 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: GenBank: 20330981 |
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| #13: RNA chain | Mass: 34404.438 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) |
| #14: RNA chain | Mass: 87892.727 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) unidentified adenovirus |
| #15: RNA chain | Mass: 60186.445 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: GenBank: 340097 |
-U5 small nuclear ribonucleoprotein ... , 2 types, 2 molecules DE
| #4: Protein | Mass: 244823.422 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O75643, RNA helicase |
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| #5: Protein | Mass: 39359.492 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q96DI7 |
-Small nuclear ribonucleoprotein ... , 6 types, 12 molecules ahcjdkfmelgn
| #6: Protein | Mass: 13940.308 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62318#8: Protein | Mass: 13310.653 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62314#9: Protein | Mass: 13551.928 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62316#10: Protein | Mass: 9734.171 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62306#11: Protein | Mass: 10817.601 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62304#12: Protein | Mass: 8508.084 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62308 |
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-U2 small nuclear ribonucleoprotein ... , 2 types, 2 molecules op
| #16: Protein | Mass: 28456.584 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P09661 |
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| #17: Protein | Mass: 25524.367 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P08579 |
-Splicing factor 3A subunit ... , 3 types, 3 molecules wuv
| #18: Protein | Mass: 58934.844 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q12874, UniProt: Q15459*PLUS |
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| #19: Protein | Mass: 88991.094 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q15459, UniProt: Q15428*PLUS |
| #20: Protein | Mass: 49327.355 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q15428, UniProt: Q12874*PLUS |
-Splicing factor 3B subunit ... , 6 types, 6 molecules 123457
| #21: Protein | Mass: 146024.938 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O75533 |
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| #22: Protein | Mass: 100377.812 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q13435 |
| #23: Protein | Mass: 135718.844 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q15393 |
| #24: Protein | Mass: 44436.570 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q15427 |
| #25: Protein | Mass: 14606.900 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9Y3B4 |
| #27: Protein | Mass: 10149.369 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9BWJ5 |
-Pre-mRNA-splicing factor ... , 2 types, 2 molecules Vx
| #34: Protein | Mass: 105646.578 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9HCG8 |
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| #39: Protein | Mass: 119443.633 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O60231, RNA helicase |
-Non-polymers , 4 types, 12 molecules 






| #40: Chemical | ChemComp-IHP / | ||
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| #41: Chemical | ChemComp-GTP / | ||
| #42: Chemical | ChemComp-MG / #43: Chemical | ChemComp-ZN / |
-Details
| Has protein modification | Y |
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-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: early Bact spliceosome / Type: COMPLEX / Entity ID: #1-#39 / Source: NATURAL |
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| Source (natural) | Organism: Homo sapiens (human) |
| Buffer solution | pH: 7.9 / Details: 20 mM HEPES-KOH, pH 7.9, 150 mM NaCl, 1.5 mM MgCl2 |
| 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: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Alignment procedure: BASIC |
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
| Image recording | Electron dose: 48 e/Å2 / Detector mode: SUPER-RESOLUTION / Film or detector model: GATAN K2 SUMMIT (4k x 4k) |
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Processing
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| CTF correction | Type: NONE | ||||||||||||||||||||||||
| Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||
| 3D reconstruction | Resolution: 4.9 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 96523 / Algorithm: FOURIER SPACE / Symmetry type: POINT | ||||||||||||||||||||||||
| Refinement | Highest resolution: 4.9 Å |
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About Yorodumi



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












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