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Yorodumi- PDB-8ro1: Structure of the C. elegans Intron Lariat Spliceosome double-prim... -
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Open data
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Basic information
| Entry | Database: PDB / ID: 8ro1 | ||||||
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| Title | Structure of the C. elegans Intron Lariat Spliceosome double-primed for disassembly (ILS'') | ||||||
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Keywords | SPLICING / pre-mRNA splicing / intron lariat spliceosome / gene expression / spliceosome | ||||||
| Function / homology | Function and homology informationfeminization of hermaphroditic germ-line / molting cycle / regulation of primary miRNA processing / SLBP independent Processing of Histone Pre-mRNAs / snRNP Assembly / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / Transport of Mature mRNA derived from an Intron-Containing Transcript / RNA lariat debranching enzyme activator activity / mRNA Splicing - Minor Pathway / Formation of TC-NER Pre-Incision Complex ...feminization of hermaphroditic germ-line / molting cycle / regulation of primary miRNA processing / SLBP independent Processing of Histone Pre-mRNAs / snRNP Assembly / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / Transport of Mature mRNA derived from an Intron-Containing Transcript / RNA lariat debranching enzyme activator activity / mRNA Splicing - Minor Pathway / Formation of TC-NER Pre-Incision Complex / Dual incision in TC-NER / Gap-filling DNA repair synthesis and ligation in TC-NER / Downregulation of SMAD2/3:SMAD4 transcriptional activity / mRNA Splicing - Major Pathway / germline cell cycle switching, mitotic to meiotic cell cycle / RNA Polymerase II Transcription Termination / mRNA Polyadenylation / vulval development / nematode larval development / egg-laying behavior / spliceosomal complex disassembly / post-spliceosomal complex / U2-type post-mRNA release spliceosomal complex / spliceosomal conformational changes to generate catalytic conformation / uterus development / post-mRNA release spliceosomal complex / apoptotic DNA fragmentation / generation of catalytic spliceosome for first transesterification step / nuclear mRNA surveillance / nuclease activity / germ cell development / embryo development ending in birth or egg hatching / U12-type spliceosomal complex / pICln-Sm protein complex / U2-type catalytic step 1 spliceosome / RNA splicing, via transesterification reactions / locomotion / SMN-Sm protein complex / spliceosomal tri-snRNP complex / commitment complex / P granule / U4 snRNP / snRNP binding / mRNA cis splicing, via spliceosome / U2-type catalytic step 2 spliceosome / U2-type spliceosomal complex / pre-mRNA binding / U1 snRNP / U2 snRNP / U2-type prespliceosome / cyclosporin A binding / generation of catalytic spliceosome for second transesterification step / precatalytic spliceosome / mRNA 3'-splice site recognition / Prp19 complex / spliceosomal tri-snRNP complex assembly / U5 snRNP / U5 snRNA binding / pre-mRNA intronic binding / U2 snRNA binding / U6 snRNA binding / U4/U6 x U5 tri-snRNP complex / U1 snRNA binding / protein K63-linked ubiquitination / spliceosomal snRNP assembly / catalytic step 2 spliceosome / spliceosomal complex / RNA splicing / helicase activity / antiviral innate immune response / RNA polymerase II transcription regulatory region sequence-specific DNA binding / peptidylprolyl isomerase / peptidyl-prolyl cis-trans isomerase activity / mRNA splicing, via spliceosome / RING-type E3 ubiquitin transferase / mRNA processing / metallopeptidase activity / ubiquitin-protein transferase activity / rRNA processing / regulation of gene expression / ubiquitin protein ligase activity / protein folding / nucleic acid binding / DNA-binding transcription factor activity, RNA polymerase II-specific / cell differentiation / RNA helicase activity / RNA helicase / DNA repair / apoptotic process / mRNA binding / GTPase activity / regulation of transcription by RNA polymerase II / positive regulation of DNA-templated transcription / GTP binding / ATP hydrolysis activity / DNA binding / RNA binding / nucleoplasm / zinc ion binding / ATP binding Similarity search - Function | ||||||
| Biological species | ![]() | ||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3 Å | ||||||
Authors | Vorlaender, M.K. / Rothe, P. / Plaschka, C. | ||||||
| Funding support | European Union, 1items
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Citation | Journal: Nature / Year: 2024Title: Mechanism for the initiation of spliceosome disassembly. Authors: Matthias K Vorländer / Patricia Rothe / Justus Kleifeld / Eric D Cormack / Lalitha Veleti / Daria Riabov-Bassat / Laura Fin / Alex W Phillips / Luisa Cochella / Clemens Plaschka / ![]() Abstract: Precursor-mRNA (pre-mRNA) splicing requires the assembly, remodelling and disassembly of the multi-megadalton ribonucleoprotein complex called the spliceosome. Recent studies have shed light on ...Precursor-mRNA (pre-mRNA) splicing requires the assembly, remodelling and disassembly of the multi-megadalton ribonucleoprotein complex called the spliceosome. Recent studies have shed light on spliceosome assembly and remodelling for catalysis, but the mechanism of disassembly remains unclear. Here we report cryo-electron microscopy structures of nematode and human terminal intron lariat spliceosomes along with biochemical and genetic data. Our results uncover how four disassembly factors and the conserved RNA helicase DHX15 initiate spliceosome disassembly. The disassembly factors probe large inner and outer spliceosome surfaces to detect the release of ligated mRNA. Two of these factors, TFIP11 and C19L1, and three general spliceosome subunits, SYF1, SYF2 and SDE2, then dock and activate DHX15 on the catalytic U6 snRNA to initiate disassembly. U6 therefore controls both the start and end of pre-mRNA splicing. Taken together, our results explain the molecular basis of the initiation of canonical spliceosome disassembly and provide a framework to understand general spliceosomal RNA helicase control and the discard of aberrant spliceosomes. | ||||||
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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 | 8ro1.cif.gz | 2.9 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb8ro1.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 8ro1.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/ro/8ro1 ftp://data.pdbj.org/pub/pdb/validation_reports/ro/8ro1 | HTTPS FTP |
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-Related structure data
| Related structure data | ![]() 19398MC ![]() 8ro0C ![]() 8ro2C ![]() 9fmdC C: citing same article ( M: map data used to model this data |
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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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| 1 |
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Components
-RNA chain , 4 types, 4 molecules 256IN
| #1: RNA chain | Mass: 68422.445 Da / Num. of mol.: 1 / Source method: isolated from a natural source Details: Full sequence: AUCGCUUCUUCGGCUUAUUAGCUAAGAUCAAAGUGUAGUAUCUGUUCUUAUCGUAUUAACCUACGGUAUACACUCGAAUGAGUGUAAUAAAGGUUAUAUGAUUUUUGGAACCUAGGGAAGACUCGGGGCUUGCUCCGACUUCCCAAGGGUCGUCCUGGCGUUGCACUGCUGCCGGGCUCGGCCCAGUCCCC Source: (natural) ![]() |
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| #2: RNA chain | Mass: 35826.145 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
| #3: RNA chain | Mass: 32483.355 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() |
| #10: RNA chain | Mass: 10348.173 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
-Pre-mRNA-splicing factor ... , 6 types, 6 molecules ADXIKMO
| #4: Protein | Mass: 272396.156 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
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| #7: Protein | Mass: 84495.477 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
| #9: Protein | Mass: 99675.094 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
| #12: Protein | Mass: 27679.885 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
| #16: Protein | Mass: 27719.021 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
| #18: Protein | Mass: 45902.750 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
-Protein , 16 types, 20 molecules CDJLNPPXQRTFXZbiopqrst
| #5: Protein | Mass: 110612.859 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() | ||||||
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| #6: Protein | Mass: 31326.172 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() | ||||||
| #11: Protein | Mass: 88116.000 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() | ||||||
| #13: Protein | Mass: 85843.469 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() | ||||||
| #17: Protein | Mass: 17153.879 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() | ||||||
| #19: Protein | Mass: 26154.846 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() | ||||||
| #20: Protein | Mass: 94244.891 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() | ||||||
| #21: Protein | Mass: 170397.688 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() | ||||||
| #22: Protein | Mass: 60303.516 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() | ||||||
| #25: Protein | Mass: 94421.539 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() | ||||||
| #27: Protein | Mass: 57466.109 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() | ||||||
| #28: Protein | Mass: 8135.504 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() | ||||||
| #30: Protein | Mass: 16768.627 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) ![]() #36: Protein | | Mass: 28905.900 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() #37: Protein | | Mass: 24881.344 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() #38: Protein | Mass: 53272.633 Da / Num. of mol.: 4 / Source method: isolated from a natural source / Source: (natural) ![]() References: UniProt: Q10051, RING-type E3 ubiquitin transferase |
-WD REPEATS REGION domain-containing ... , 3 types, 3 molecules ETW
| #8: Protein | Mass: 36865.559 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
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| #24: Protein | Mass: 54766.215 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
| #26: Protein | Mass: 65385.664 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
-CWF19-like protein ... , 2 types, 2 molecules L1L2
| #14: Protein | Mass: 59034.898 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
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| #15: Protein | Mass: 53269.211 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
-Peptidyl-prolyl cis-trans ... , 2 types, 2 molecules Sy
| #23: Protein | Mass: 18547.002 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
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| #39: Protein | Mass: 8825.046 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
-Small nuclear ribonucleoprotein Sm ... , 2 types, 4 molecules ahcj
| #29: Protein | Mass: 14836.212 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) ![]() #31: Protein | Mass: 13724.070 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) ![]() |
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-Probable small nuclear ribonucleoprotein ... , 4 types, 8 molecules dkelfmgn
| #32: Protein | Mass: 13291.529 Da / Num. of mol.: 2 / Source method: isolated from a natural source Details: MSAQAKPRSEMTAEELAAKEDEEFNVGPLSILTNSVKNNHQVLINCRNNKKLLGRVKAFDRHCNMVLENVKEMWTEVPKTGKGKKKAKSVAKDRFISKMFLRGDSVILVVKNPLAQAE Source: (natural) ![]() #33: Protein | Mass: 10625.318 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) ![]() #34: Protein | Mass: 9256.534 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) ![]() #35: Protein | Mass: 8756.209 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) ![]() |
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-Non-polymers , 4 types, 17 molecules 






| #40: Chemical | ChemComp-MG / #41: Chemical | #42: Chemical | ChemComp-GTP / | #43: Chemical | ChemComp-ZN / |
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-Details
| Has ligand of interest | N |
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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: Intron lariat spliceosome (ILS'') / Type: COMPLEX / Entity ID: #1-#39 / Source: NATURAL |
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| Source (natural) | Organism: ![]() |
| Buffer solution | pH: 7.9 |
| 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 / Nominal defocus max: 2000 nm / Nominal defocus min: 750 nm |
| Image recording | Electron dose: 60 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
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Processing
| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
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| 3D reconstruction | Resolution: 3 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 247908 / Symmetry type: POINT |
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