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Yorodumi- PDB-5xjc: Cryo-EM structure of the human spliceosome just prior to exon lig... -
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Basic information
| Entry | Database: PDB / ID: 5xjc | |||||||||
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| Title | Cryo-EM structure of the human spliceosome just prior to exon ligation at 3.6 angstrom | |||||||||
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Keywords | SPLICING / RNA splicing / human spliceosome / C* complex / atomic structure / step 2 factors / EJC | |||||||||
| Function / homology | Function and homology informationsecond spliceosomal transesterification activity / regulation of RNA helicase activity / exon-exon junction subcomplex mago-y14 / negative regulation of selenocysteine incorporation / cellular response to selenite ion / renal system process / regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / exon-exon junction complex / regulation of vitamin D receptor signaling pathway / pre-mRNA 3'-splice site binding ...second spliceosomal transesterification activity / regulation of RNA helicase activity / exon-exon junction subcomplex mago-y14 / negative regulation of selenocysteine incorporation / cellular response to selenite ion / renal system process / regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / exon-exon junction complex / regulation of vitamin D receptor signaling pathway / pre-mRNA 3'-splice site binding / selenocysteine insertion sequence binding / mRNA metabolic process / DEAD/H-box RNA helicase binding / regulation of translation at postsynapse, modulating synaptic transmission / regulation of retinoic acid receptor signaling pathway / alternative mRNA splicing, via spliceosome / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / generation of catalytic spliceosome for first transesterification step / Z-decay: degradation of maternal mRNAs by zygotically expressed factors / 3'-5' RNA helicase activity / cis assembly of pre-catalytic spliceosome / histone pre-mRNA 3'end processing complex / nuclear histone mRNA catabolic process / embryonic cranial skeleton morphogenesis / U4atac snRNP / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / U4atac/U6atac snRNP / regulation of mRNA processing / Deadenylation of mRNA / C2H2 zinc finger domain binding / U11/U12 snRNP / negative regulation of excitatory postsynaptic potential / poly(A) binding / U4atac/U6atac x U5 tri-snRNP complex / embryonic brain development / 7-methylguanosine cap hypermethylation / U12-type spliceosomal complex / U1 snRNP binding / regulation of mRNA splicing, via spliceosome / nuclear retinoic acid receptor binding / post-spliceosomal complex / positive regulation of mRNA splicing, via spliceosome / RNA splicing, via transesterification reactions / pICln-Sm protein complex / snRNP binding / ATP-dependent activity, acting on RNA / mRNA 3'-end processing / sno(s)RNA-containing ribonucleoprotein complex / methylosome / U12-type catalytic step 2 spliceosome / small nuclear ribonucleoprotein complex / RNA Polymerase II Transcription Termination / SMN-Sm protein complex / post-mRNA release spliceosomal complex / U12-type precatalytic spliceosome / spliceosomal tri-snRNP complex / M-decay: degradation of maternal mRNAs by maternally stored factors / positive regulation of vitamin D receptor signaling pathway / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / P granule / commitment complex / 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 / telomerase holoenzyme complex / Transport of Mature mRNA derived from an Intron-Containing Transcript / U2-type prespliceosome assembly / NOTCH4 Intracellular Domain Regulates Transcription / telomerase RNA binding / pre-mRNA binding / positive regulation of neurogenesis / U4/U6 snRNP / U2-type catalytic step 2 spliceosome / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / U1 snRNP / U2 snRNP / NOTCH3 Intracellular Domain Regulates Transcription / protein methylation / U2-type catalytic step 1 spliceosome / exploration behavior / U4 snRNP / mRNA stabilization / U2-type prespliceosome / mRNA cis splicing, via spliceosome / protein peptidyl-prolyl isomerization / nuclear androgen receptor binding / K63-linked polyubiquitin modification-dependent protein binding / ubiquitin-ubiquitin ligase activity / Notch-HLH transcription pathway / Formation of paraxial mesoderm / regulation of alternative mRNA splicing, via spliceosome / precatalytic spliceosome Similarity search - Function | |||||||||
| Biological species | Homo sapiens (human)![]() Human adenovirus 2 | |||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||
Authors | Zhang, X. / Yan, C. / Hang, J. / Finci, I.L. / Lei, J. / Shi, Y. | |||||||||
| Funding support | China, 2items
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Citation | Journal: Cell / Year: 2017Title: An Atomic Structure of the Human Spliceosome. Authors: Xiaofeng Zhang / Chuangye Yan / Jing Hang / Lorenzo I Finci / Jianlin Lei / Yigong Shi / ![]() Abstract: Mechanistic understanding of pre-mRNA splicing requires detailed structural information on various states of the spliceosome. Here we report the cryo electron microscopy (cryo-EM) structure of the ...Mechanistic understanding of pre-mRNA splicing requires detailed structural information on various states of the spliceosome. Here we report the cryo electron microscopy (cryo-EM) structure of the human spliceosome just before exon ligation (the C complex) at an average resolution of 3.76 Å. The splicing factor Prp17 stabilizes the active site conformation. The step II factor Slu7 adopts an extended conformation, binds Prp8 and Cwc22, and is poised for selection of the 3'-splice site. Remarkably, the intron lariat traverses through a positively charged central channel of RBM22; this unusual organization suggests mechanisms of intron recruitment, confinement, and release. The protein PRKRIP1 forms a 100-Å α helix linking the distant U2 snRNP to the catalytic center. A 35-residue fragment of the ATPase/helicase Prp22 latches onto Prp8, and the quaternary exon junction complex (EJC) recognizes upstream 5'-exon sequences and associates with Cwc22 and the GTPase Snu114. These structural features reveal important mechanistic insights into exon ligation. | |||||||||
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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 | 5xjc.cif.gz | 3.1 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb5xjc.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 5xjc.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/xj/5xjc ftp://data.pdbj.org/pub/pdb/validation_reports/xj/5xjc | HTTPS FTP |
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-Related structure data
| Related structure data | ![]() 6721MC M: map data used to model this data C: citing same article ( |
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| Similar structure data |
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Assembly
| Deposited unit | ![]()
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Components
-Protein , 18 types, 19 molecules ACJLNPQRSTUXYbiuvwx
| #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 | ||||||||
| #10: Protein | Mass: 100610.008 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9BZJ0 | ||||||||
| #12: Protein | Mass: 92406.883 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q99459 | ||||||||
| #14: Protein | Mass: 17032.850 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P41223 | ||||||||
| #16: Protein | Mass: 26674.447 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9P013 | ||||||||
| #17: Protein | Mass: 171502.453 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O60306 | ||||||||
| #18: Protein | Mass: 61770.648 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q13573 | ||||||||
| #19: Protein | Mass: 18257.805 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9Y3C6, peptidylprolyl isomerase | ||||||||
| #20: Protein | Mass: 57280.758 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O43660 | ||||||||
| #21: Protein | Mass: 300255.312 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9UQ35 | ||||||||
| #24: Protein | Mass: 21040.176 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9H875 | ||||||||
| #25: Protein | Mass: 139508.328 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q14562, RNA helicase | ||||||||
| #28: Protein | Mass: 23686.004 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P14678#37: Protein | | Mass: 46930.961 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P38919, RNA helicase#38: Protein | | Mass: 17301.799 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q96A72#39: Protein | | Mass: 19925.070 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9Y5S9#40: Protein | | Mass: 76381.992 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O15234 |
-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) |
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| #6: RNA chain | Mass: 34404.438 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) |
| #7: RNA chain | Mass: 87977.836 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) ![]() Human adenovirus 2 |
| #8: RNA chain | Mass: 60186.445 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: GenBank: 609501 |
-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 |
-Pre-mRNA-splicing factor ... , 6 types, 6 molecules IKMOVZ
| #9: Protein | Mass: 100148.711 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9HCS7 |
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| #11: Protein | Mass: 26163.420 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O75934 |
| #13: Protein | Mass: 28780.518 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O95926 |
| #15: Protein | Mass: 46959.555 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9NW64 |
| #22: Protein | Mass: 105646.578 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9HCG8 |
| #26: Protein | Mass: 68510.844 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O95391 |
-Pre-mRNA-processing factor ... , 2 types, 5 molecules Wqrst
| #23: Protein | Mass: 65612.180 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O60508 |
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| #36: Protein | Mass: 55245.547 Da / Num. of mol.: 4 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human)References: UniProt: Q9UMS4, RING-type E3 ubiquitin transferase |
-Small nuclear ribonucleoprotein ... , 6 types, 12 molecules ahcjdkfmelgn
| #27: Protein | Mass: 13940.308 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62318#29: Protein | Mass: 13310.653 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62314#30: Protein | Mass: 13551.928 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62316#31: Protein | Mass: 9734.171 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62306#32: Protein | Mass: 10817.601 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62304#33: 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
| #34: 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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| #35: Protein | Mass: 25524.367 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P08579 |
-Non-polymers , 6 types, 24 molecules 










| #41: Chemical | ChemComp-IHP / | ||||||
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| #42: Chemical | ChemComp-GTP / | ||||||
| #43: Chemical | ChemComp-MG / #44: Chemical | #45: Chemical | ChemComp-ZN / #46: Chemical | |
-Details
| Has protein modification | Y |
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| Sequence details | The UNK reisue between G-1 and G1 is added intentionally as per authors request to fill in the ...The UNK reisue between G-1 and G1 is added intentionally as per authors request to fill in the cleavage by RNA splicing. |
-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: human C* spliceosome / Type: COMPLEX / Entity ID: #1-#40 / Source: NATURAL |
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| Source (natural) | Organism: Homo sapiens (human) |
| Buffer solution | pH: 7.5 |
| 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 / Cs: 2.7 mm / Alignment procedure: BASIC |
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: GATAN LIQUID NITROGEN |
| Image recording | Average exposure time: 0.25 sec. / Electron dose: 50 e/Å2 / Detector mode: SUPER-RESOLUTION / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Num. of grids imaged: 5 / Num. of real images: 7308 |
| EM imaging optics | Energyfilter name: GIF Quantum LS / Energyfilter upper: 10 eV / Energyfilter lower: 0 eV / Chromatic aberration corrector: -10 |
| Image scans | Movie frames/image: 32 / Used frames/image: 2-32 |
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Processing
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||
| 3D reconstruction | Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 100085 / Symmetry type: POINT | ||||||||||||||||
| Refinement | Highest resolution: 3.6 Å |
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About Yorodumi



Homo sapiens (human)
Human adenovirus 2
China, 2items
Citation
UCSF Chimera









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