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Open data
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Basic information
| Entry | Database: PDB / ID: 7evo | |||||||||||||||||||||||||||||||||
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| Title | The cryo-EM structure of the human 17S U2 snRNP | |||||||||||||||||||||||||||||||||
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Keywords | SPLICING / U2 snRNP / PRP5 / SF3B1 | |||||||||||||||||||||||||||||||||
| Function / homology | Function and homology informationU11/U12 snRNP / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / histone pre-mRNA 3'end processing complex / nuclear histone mRNA catabolic process / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / poly-ADP-D-ribose modification-dependent protein binding ...U11/U12 snRNP / U2 snRNP binding / U7 snRNA binding / histone pre-mRNA DCP binding / U7 snRNP / histone pre-mRNA 3'end processing complex / nuclear histone mRNA catabolic process / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / poly-ADP-D-ribose modification-dependent protein binding / 7-methylguanosine cap hypermethylation / U12-type spliceosomal complex / U1 snRNP binding / pICln-Sm protein complex / methylosome / RNA splicing, via transesterification reactions / small nuclear ribonucleoprotein complex / splicing factor binding / SMN-Sm protein complex / protein localization to site of double-strand break / spliceosomal tri-snRNP complex / commitment complex / P granule / U4 snRNP / snRNP binding / U2-type precatalytic spliceosome / mRNA cis splicing, via spliceosome / U2-type prespliceosome assembly / U2-type catalytic step 2 spliceosome / U2-type spliceosomal complex / telomerase holoenzyme complex / telomerase RNA binding / pre-mRNA binding / U1 snRNP / U2 snRNP / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / chromatin-protein adaptor activity / RNA Polymerase II Transcription Termination / protein methylation / U2-type prespliceosome / precatalytic spliceosome / mRNA 3'-splice site recognition / regulation of RNA splicing / positive regulation of transcription by RNA polymerase III / mRNA Splicing - Minor Pathway / positive regulation of transcription by RNA polymerase I / spliceosomal complex assembly / negative regulation of mRNA splicing, via spliceosome / spliceosomal tri-snRNP complex assembly / U5 snRNP / U2 snRNA binding / U4/U6 x U5 tri-snRNP complex / Cajal body / U1 snRNA binding / spliceosomal snRNP assembly / stem cell differentiation / catalytic step 2 spliceosome / spliceosomal complex / mRNA Splicing - Major Pathway / Gene and protein expression by JAK-STAT signaling after Interleukin-12 stimulation / RNA splicing / regulation of DNA repair / positive regulation of neuron projection development / negative regulation of protein catabolic process / mRNA splicing, via spliceosome / double-strand break repair via homologous recombination / B-WICH complex positively regulates rRNA expression / mRNA processing / mRNA Polyadenylation / nuclear matrix / site of double-strand break / Dengue Virus-Host Interactions / snRNP Assembly / spermatogenesis / SARS-CoV-2 modulates host translation machinery / nuclear body / RNA helicase activity / nuclear speck / RNA helicase / chromatin remodeling / hydrolase activity / mRNA binding / nucleolus / positive regulation of DNA-templated transcription / protein-containing complex binding / chromatin / enzyme binding / positive regulation of transcription by RNA polymerase II / DNA binding / DNA-templated transcription / RNA binding / extracellular exosome / nucleoplasm / zinc ion binding / ATP binding / nucleus / cytosol / cytoplasm Similarity search - Function | |||||||||||||||||||||||||||||||||
| Biological species | Homo sapiens (human) | |||||||||||||||||||||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.5 Å | |||||||||||||||||||||||||||||||||
Authors | Zhang, X. / Zhan, X. / Shi, Y. | |||||||||||||||||||||||||||||||||
| Funding support | China, 1items
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Citation | Journal: Nat Struct Mol Biol / Year: 2024Title: Structural insights into branch site proofreading by human spliceosome. Authors: Xiaofeng Zhang / Xiechao Zhan / Tong Bian / Fenghua Yang / Pan Li / Yichen Lu / Zhihan Xing / Rongyan Fan / Qiangfeng Cliff Zhang / Yigong Shi / ![]() Abstract: Selection of the pre-mRNA branch site (BS) by the U2 small nuclear ribonucleoprotein (snRNP) is crucial to prespliceosome (A complex) assembly. The RNA helicase PRP5 proofreads BS selection but the ...Selection of the pre-mRNA branch site (BS) by the U2 small nuclear ribonucleoprotein (snRNP) is crucial to prespliceosome (A complex) assembly. The RNA helicase PRP5 proofreads BS selection but the underlying mechanism remains unclear. Here we report the atomic structures of two sequential complexes leading to prespliceosome assembly: human 17S U2 snRNP and a cross-exon pre-A complex. PRP5 is anchored on 17S U2 snRNP mainly through occupation of the RNA path of SF3B1 by an acidic loop of PRP5; the helicase domain of PRP5 associates with U2 snRNA; the BS-interacting stem-loop (BSL) of U2 snRNA is shielded by TAT-SF1, unable to engage the BS. In the pre-A complex, an initial U2-BS duplex is formed; the translocated helicase domain of PRP5 stays with U2 snRNA and the acidic loop still occupies the RNA path. The pre-A conformation is specifically stabilized by the splicing factors SF1, DNAJC8 and SF3A2. Cancer-derived mutations in SF3B1 damage its association with PRP5, compromising BS proofreading. Together, these findings reveal key insights into prespliceosome assembly and BS selection or proofreading by PRP5. | |||||||||||||||||||||||||||||||||
| History |
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Structure visualization
| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 7evo.cif.gz | 911.4 KB | Display | PDBx/mmCIF format |
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| PDB format | pdb7evo.ent.gz | 626.6 KB | Display | PDB format |
| PDBx/mmJSON format | 7evo.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/ev/7evo ftp://data.pdbj.org/pub/pdb/validation_reports/ev/7evo | HTTPS FTP |
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-Related structure data
| Related structure data | ![]() 31334MC ![]() 7vpxC M: map data used to model this data C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
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Assembly
| Deposited unit | ![]()
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Components
-RNA chain , 1 types, 1 molecules H
| #1: 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 |
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-Splicing factor 3B subunit ... , 5 types, 5 molecules 12345
| #2: 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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| #3: Protein | Mass: 100377.812 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q13435 |
| #4: Protein | Mass: 135718.844 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q15393 |
| #5: Protein | Mass: 44436.570 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q15427 |
| #6: Protein | Mass: 10149.369 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9BWJ5 |
-Protein , 4 types, 4 molecules 6DEf
| #7: Protein | Mass: 12427.524 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q7RTV0 |
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| #11: Protein | Mass: 85965.961 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: O43719 |
| #12: Protein | Mass: 119807.664 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: DDX46 / Cell line (production host): HEK293 / Production host: Homo sapiens (human) / References: UniProt: A0A0C4DG89, RNA helicase |
| #20: Protein | Mass: 24642.131 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P14678 |
-Splicing factor 3A subunit ... , 3 types, 3 molecules ABC
| #8: Protein | Mass: 88991.094 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q15459 |
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| #9: Protein | Mass: 49327.355 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q15428 |
| #10: Protein | Mass: 58934.844 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q12874 |
-U2 small nuclear ribonucleoprotein ... , 2 types, 2 molecules FG
| #13: 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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| #14: Protein | Mass: 25524.367 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P08579 |
-Small nuclear ribonucleoprotein ... , 6 types, 6 molecules abcdeg
| #15: Protein | Mass: 13551.928 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62316 |
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| #16: Protein | Mass: 9734.171 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62306 |
| #17: Protein | Mass: 10817.601 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62304 |
| #18: Protein | Mass: 8508.084 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62308 |
| #19: Protein | Mass: 13940.308 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62318 |
| #21: Protein | Mass: 13310.653 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P62314 |
-Non-polymers , 2 types, 5 molecules 


| #22: Chemical | ChemComp-9B0 / [( |
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| #23: Chemical | ChemComp-ZN / |
-Details
| Has ligand of interest | Y |
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| Has protein modification | N |
-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: The human U2 snRNP / Type: COMPLEX / Entity ID: #1-#21 / Source: NATURAL |
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| Molecular weight | Experimental value: NO |
| Source (natural) | Organism: Homo sapiens (human) |
| Buffer solution | pH: 7.9 |
| Specimen | Conc.: 0.5 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Specimen support | Grid material: GOLD / Grid type: Quantifoil R1.2/1.3 |
| Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % |
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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 |
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
| Image recording | Electron dose: 50 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
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Processing
| Software | Name: PHENIX / Version: 1.17.1_3660: / Classification: refinement | ||||||||||||||||||||||||
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| CTF correction | Type: NONE | ||||||||||||||||||||||||
| 3D reconstruction | Resolution: 2.5 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 485418 / Algorithm: FOURIER SPACE / Symmetry type: POINT | ||||||||||||||||||||||||
| Refine LS restraints |
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About Yorodumi




Homo sapiens (human)
China, 1items
Citation


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FIELD EMISSION GUN