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Yorodumi- PDB-9fmd: Integrative model of the human post-catalytic spliceosome (P-complex) -
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Open data
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Basic information
| Entry | Database: PDB / ID: 9fmd | |||||||||
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| Title | Integrative model of the human post-catalytic spliceosome (P-complex) | |||||||||
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Keywords | SPLICING / P-complex / spliceosome | |||||||||
| Function / homology | Function and homology informationregulation of homologous chromosome segregation / second spliceosomal transesterification activity / endoplasmic reticulum membrane organization / exon-exon junction subcomplex mago-y14 / negative regulation of selenocysteine incorporation / negative regulation of nitric-oxide synthase activity / cellular response to selenite ion / post-spliceosomal complex / renal system process / exon-exon junction complex ...regulation of homologous chromosome segregation / second spliceosomal transesterification activity / endoplasmic reticulum membrane organization / exon-exon junction subcomplex mago-y14 / negative regulation of selenocysteine incorporation / negative regulation of nitric-oxide synthase activity / cellular response to selenite ion / post-spliceosomal complex / renal system process / exon-exon junction complex / regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / regulation of vitamin D receptor signaling pathway / pre-mRNA 3'-splice site binding / selenocysteine insertion sequence binding / mRNA metabolic process / NOSIP mediated eNOS trafficking / protein exit from endoplasmic reticulum / regulation of translation at postsynapse, modulating synaptic transmission / regulation of retinoic acid receptor signaling pathway / alternative mRNA splicing, via spliceosome / negative regulation of toll-like receptor signaling pathway / post-mRNA release spliceosomal complex / U2 snRNP binding / negative regulation of catalytic activity / U7 snRNA binding / histone pre-mRNA DCP binding / generation of catalytic spliceosome for first transesterification step / U7 snRNP / 3'-5' RNA helicase activity / Z-decay: degradation of maternal mRNAs by zygotically expressed factors / cis assembly of pre-catalytic spliceosome / histone pre-mRNA 3'end processing complex / nuclear histone mRNA catabolic process / negative regulation of lipopolysaccharide-mediated signaling pathway / embryonic cranial skeleton morphogenesis / endonucleolytic cleavage of tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / regulation of mRNA processing / SLBP independent Processing of Histone Pre-mRNAs / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / Deadenylation of mRNA / negative regulation of interleukin-8 production / negative regulation of excitatory postsynaptic potential / negative regulation of interferon-beta production / positive regulation of alpha-beta T cell differentiation / embryonic brain development / poly(A) binding / 7-methylguanosine cap hypermethylation / U12-type spliceosomal complex / positive regulation of mRNA splicing, via spliceosome / nuclear retinoic acid receptor binding / U1 snRNP binding / C2H2 zinc finger domain binding / pICln-Sm protein complex / regulation of mRNA splicing, via spliceosome / U2-type catalytic step 1 spliceosome / methylosome / ATP-dependent activity, acting on RNA / RNA splicing, via transesterification reactions / mRNA 3'-end processing / sno(s)RNA-containing ribonucleoprotein complex / small nuclear ribonucleoprotein complex / M-decay: degradation of maternal mRNAs by maternally stored factors / SMN-Sm protein complex / spliceosomal tri-snRNP complex / regulation of nitric oxide biosynthetic process / positive regulation of vitamin D receptor signaling pathway / commitment complex / P granule / U4 snRNP / snRNP binding / 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 / U2-type prespliceosome assembly / RUNX3 regulates NOTCH signaling / U2-type catalytic step 2 spliceosome / Notch binding / U2-type spliceosomal complex / exploration behavior / nuclear vitamin D receptor binding / telomerase holoenzyme complex / Transport of Mature mRNA derived from an Intron-Containing Transcript / NOTCH4 Intracellular Domain Regulates Transcription / telomerase RNA binding / pre-mRNA binding / positive regulation of neurogenesis / U1 snRNP / U2 snRNP / spliceosome conformational change to release U4 (or U4atac) and U1 (or U11) / negative regulation of type I interferon-mediated signaling pathway / RNA Polymerase II Transcription Termination / NOTCH3 Intracellular Domain Regulates Transcription / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / protein methylation / mRNA stabilization / U2-type prespliceosome / protein peptidyl-prolyl isomerization / K63-linked polyubiquitin modification-dependent protein binding / nuclear androgen receptor binding Similarity search - Function | |||||||||
| Biological species | Homo sapiens (human)![]() Human adenovirus 2 | |||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.3 Å | |||||||||
Authors | Rothe, P. / Plaschka, C. / Vorlaender, M.K. | |||||||||
| 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. #1: Journal: Science / Year: 2019Title: A human postcatalytic spliceosome structure reveals essential roles of metazoan factors for exon ligation. Authors: Sebastian M Fica / Chris Oubridge / Max E Wilkinson / Andrew J Newman / Kiyoshi Nagai / ![]() Abstract: During exon ligation, the spliceosome recognizes the 3'-splice site (3'SS) of precursor messenger RNA (pre-mRNA) through non-Watson-Crick pairing with the 5'SS and the branch adenosine, in a ...During exon ligation, the spliceosome recognizes the 3'-splice site (3'SS) of precursor messenger RNA (pre-mRNA) through non-Watson-Crick pairing with the 5'SS and the branch adenosine, in a conformation stabilized by Prp18 and Prp8. Here we present the 3.3-angstrom cryo-electron microscopy structure of a human postcatalytic spliceosome just after exon ligation. The 3'SS docks at the active site through conserved RNA interactions in the absence of Prp18. Unexpectedly, the metazoan-specific FAM32A directly bridges the 5'-exon and intron 3'SS of pre-mRNA and promotes exon ligation, as shown by functional assays. CACTIN, SDE2, and NKAP-factors implicated in alternative splicing-further stabilize the catalytic conformation of the spliceosome during exon ligation. Together these four proteins act as exon ligation factors. Our study reveals how the human spliceosome has co-opted additional proteins to modulate a conserved RNA-based mechanism for 3'SS selection and to potentially fine-tune alternative splicing at the exon ligation stage. | |||||||||
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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 | 9fmd.cif.gz | 3.2 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb9fmd.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 9fmd.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/fm/9fmd ftp://data.pdbj.org/pub/pdb/validation_reports/fm/9fmd | HTTPS FTP |
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-Related structure data
| Related structure data | ![]() 4525M ![]() 8ro0C ![]() 8ro1C ![]() 8ro2C 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
+Protein , 24 types, 25 molecules 1325056789ACJLNNOPQRSRTVXZbiwz
+RNA chain , 5 types, 5 molecules 256EXIN
+Splicing factor ... , 3 types, 3 molecules 3Fx
+U5 small nuclear ribonucleoprotein ... , 2 types, 2 molecules BE
+Pre-mRNA-splicing factor ... , 7 types, 7 molecules DHIKMOU
+Peptidyl-prolyl cis-trans ... , 2 types, 2 molecules Sy
+Pre-mRNA-processing factor ... , 2 types, 5 molecules Wqrst
+Small nuclear ribonucleoprotein ... , 6 types, 12 molecules ahcjdkelfmgn
+U2 small nuclear ribonucleoprotein ... , 2 types, 2 molecules op
+Non-polymers , 6 types, 16 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
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| Molecular weight | Experimental value: NO | ||||||||||||||||||||||||||||||
| Source (natural) |
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| 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: 3500 nm / Nominal defocus min: 1500 nm |
| Image recording | Electron dose: 53 e/Å2 / Film or detector model: GATAN K2 SUMMIT (4k x 4k) |
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Processing
| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
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| 3D reconstruction | Resolution: 3.3 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 103860 / Symmetry type: POINT |
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About Yorodumi



Homo sapiens (human)
Human adenovirus 2
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FIELD EMISSION GUN