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Basic information
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Title | The cryo-EM structure of human pre-C*-II complex | |||||||||
![]() | The cryo-EM map of human pre-C*-II complex | |||||||||
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![]() | RNA splicing / PRP22 / exon ligation / human spliceosome / C* complex / SPLICING | |||||||||
Function / homology | ![]() second spliceosomal transesterification activity / exon-exon junction subcomplex mago-y14 / negative regulation of selenocysteine incorporation / regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / cellular response to selenite ion / selenocysteine insertion sequence binding / exon-exon junction complex / pre-mRNA 3'-splice site binding / negative regulation of proteasomal protein catabolic process / regulation of translation at postsynapse, modulating synaptic transmission ...second spliceosomal transesterification activity / exon-exon junction subcomplex mago-y14 / negative regulation of selenocysteine incorporation / regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / cellular response to selenite ion / selenocysteine insertion sequence binding / exon-exon junction complex / pre-mRNA 3'-splice site binding / negative regulation of proteasomal protein catabolic process / regulation of translation at postsynapse, modulating synaptic transmission / regulation of retinoic acid receptor signaling pathway / post-mRNA release spliceosomal complex / renal system process / intracellular mRNA localization / U2 snRNP binding / U7 snRNA binding / negative regulation of toll-like receptor signaling pathway / histone pre-mRNA DCP binding / 3'-5' RNA helicase activity / U7 snRNP / generation of catalytic spliceosome for first transesterification step / Z-decay: degradation of maternal mRNAs by zygotically expressed factors / histone pre-mRNA 3'end processing complex / regulation of mRNA processing / regulation of vitamin D receptor signaling pathway / negative regulation of excitatory postsynaptic potential / negative regulation of interleukin-8 production / SLBP independent Processing of Histone Pre-mRNAs / negative regulation of lipopolysaccharide-mediated signaling pathway / SLBP Dependent Processing of Replication-Dependent Histone Pre-mRNAs / Deadenylation of mRNA / nuclear retinoic acid receptor binding / embryonic brain development / protein methylation / oocyte development / embryonic cranial skeleton morphogenesis / U12-type spliceosomal complex / alternative mRNA splicing, via spliceosome / poly(A) binding / 7-methylguanosine cap hypermethylation / negative regulation of interferon-beta production / U1 snRNP binding / sno(s)RNA-containing ribonucleoprotein complex / methylosome / mRNA 3'-end processing / M-decay: degradation of maternal mRNAs by maternally stored factors / RNA splicing, via transesterification reactions / ATP-dependent activity, acting on RNA / pICln-Sm protein complex / C2H2 zinc finger domain binding / U2-type catalytic step 1 spliceosome / pre-mRNA binding / regulation of mRNA splicing, via spliceosome / snRNP binding / positive regulation of mRNA splicing, via spliceosome / small nuclear ribonucleoprotein complex / SMN-Sm protein complex / spliceosomal tri-snRNP complex / host-mediated activation of viral transcription / P granule / telomerase holoenzyme complex / U2-type precatalytic spliceosome / positive regulation of vitamin D receptor signaling pathway / commitment complex / telomerase RNA binding / mRNA cis splicing, via spliceosome / U2-type spliceosomal complex / nuclear vitamin D receptor binding / U2-type prespliceosome assembly / Notch binding / Transport of Mature mRNA derived from an Intron-Containing Transcript / Regulation of gene expression in late stage (branching morphogenesis) pancreatic bud precursor cells / U2-type catalytic step 2 spliceosome / RUNX3 regulates NOTCH signaling / positive regulation of neurogenesis / NOTCH4 Intracellular Domain Regulates Transcription / U4 snRNP / RNA Polymerase II Transcription Termination / nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / U2 snRNP / U1 snRNP / NOTCH3 Intracellular Domain Regulates Transcription / negative regulation of type I interferon-mediated signaling pathway / U2-type prespliceosome / inner cell mass cell proliferation / protein peptidyl-prolyl isomerization / WD40-repeat domain binding / ubiquitin-ubiquitin ligase activity / K63-linked polyubiquitin modification-dependent protein binding / nuclear androgen receptor binding / precatalytic spliceosome / negative regulation of NF-kappaB transcription factor activity / cyclosporin A binding / regulation of alternative mRNA splicing, via spliceosome / Notch-HLH transcription pathway / lipid biosynthetic process / Formation of paraxial mesoderm / positive regulation of transforming growth factor beta receptor signaling pathway / SMAD binding / protein kinase inhibitor activity Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||
![]() | Zhan X / Lu Y | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Mechanism of exon ligation by human spliceosome. Authors: Xiechao Zhan / Yichen Lu / Xiaofeng Zhang / Chuangye Yan / Yigong Shi / ![]() Abstract: Pre-mRNA splicing involves two sequential reactions: branching and exon ligation. The C complex after branching undergoes remodeling to become the C complex, which executes exon ligation. Here, we ...Pre-mRNA splicing involves two sequential reactions: branching and exon ligation. The C complex after branching undergoes remodeling to become the C complex, which executes exon ligation. Here, we report cryo-EM structures of two intermediate human spliceosomal complexes, pre-C-I and pre-C-II, both at 3.6 Å. In both structures, the 3' splice site is already docked into the active site, the ensuing 3' exon sequences are anchored on PRP8, and the step II factor FAM192A contacts the duplex between U2 snRNA and the branch site. In the transition of pre-C-I to pre-C-II, the step II factors Cactin, FAM32A, PRKRIP1, and SLU7 are recruited. Notably, the RNA helicase PRP22 is positioned quite differently in the pre-C-I, pre-C-II, and C complexes, suggesting a role in 3' exon binding and proofreading. Together with information on human C and C complexes, our studies recapitulate a molecular choreography of the C-to-C transition, revealing mechanistic insights into exon ligation. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 230.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 70.7 KB 70.7 KB | Display Display | ![]() |
Images | ![]() | 68.4 KB | ||
Filedesc metadata | ![]() | 21.4 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 534.1 KB | Display | ![]() |
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Full document | ![]() | 533.6 KB | Display | |
Data in XML | ![]() | 7.4 KB | Display | |
Data in CIF | ![]() | 8.6 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7w5aMC ![]() 7w59C ![]() 7w5bC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||
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Annotation | The cryo-EM map of human pre-C*-II complex | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.338 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
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Sample components
+Entire : Human pre-C*-II complex
+Supramolecule #1: Human pre-C*-II complex
+Macromolecule #1: Pre-mRNA-processing-splicing factor 8
+Macromolecule #3: 116 kDa U5 small nuclear ribonucleoprotein component
+Macromolecule #4: U5 small nuclear ribonucleoprotein 40 kDa protein
+Macromolecule #9: Pre-mRNA-splicing factor SYF1
+Macromolecule #10: Crooked neck-like protein 1
+Macromolecule #11: Pre-mRNA-splicing factor SPF27
+Macromolecule #12: Cell division cycle 5-like protein
+Macromolecule #13: Pre-mRNA-splicing factor SYF2
+Macromolecule #14: Protein BUD31 homolog
+Macromolecule #15: Pre-mRNA-splicing factor RBM22
+Macromolecule #16: Spliceosome-associated protein CWC15 homolog
+Macromolecule #17: RNA helicase aquarius
+Macromolecule #18: SNW domain-containing protein 1
+Macromolecule #19: Peptidyl-prolyl cis-trans isomerase-like 1
+Macromolecule #20: Pleiotropic regulator 1
+Macromolecule #21: Serine/arginine repetitive matrix protein 2
+Macromolecule #22: Pre-mRNA-splicing factor CWC22 homolog
+Macromolecule #23: Pre-mRNA-processing factor 17
+Macromolecule #24: PSME3-interacting protein
+Macromolecule #25: ATP-dependent RNA helicase DHX8
+Macromolecule #26: Cactin
+Macromolecule #27: PRKR-interacting protein 1
+Macromolecule #28: Protein FAM32A
+Macromolecule #29: Small nuclear ribonucleoprotein-associated proteins B and B'
+Macromolecule #30: Peptidyl-prolyl cis-trans isomerase E
+Macromolecule #31: Small nuclear ribonucleoprotein Sm D3
+Macromolecule #32: Small nuclear ribonucleoprotein Sm D1
+Macromolecule #33: Small nuclear ribonucleoprotein Sm D2
+Macromolecule #34: Small nuclear ribonucleoprotein F
+Macromolecule #35: Small nuclear ribonucleoprotein E
+Macromolecule #36: Small nuclear ribonucleoprotein G
+Macromolecule #37: Protein mago nashi homolog
+Macromolecule #38: RNA-binding protein 8A
+Macromolecule #39: Eukaryotic initiation factor 4A-III
+Macromolecule #40: Protein CASC3
+Macromolecule #41: Pre-mRNA-processing factor 19
+Macromolecule #42: U2 small nuclear ribonucleoprotein A'
+Macromolecule #43: U2 small nuclear ribonucleoprotein B''
+Macromolecule #44: Pre-mRNA-splicing factor SLU7
+Macromolecule #2: U5 snRNA
+Macromolecule #5: U6 snRNA
+Macromolecule #6: Pre-mRNA
+Macromolecule #7: Pre-mRNA
+Macromolecule #8: U2 snRNA
+Macromolecule #45: INOSITOL HEXAKISPHOSPHATE
+Macromolecule #46: GUANOSINE-5'-TRIPHOSPHATE
+Macromolecule #47: MAGNESIUM ION
+Macromolecule #48: ZINC ION
+Macromolecule #49: ADENOSINE-5'-TRIPHOSPHATE
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.9 |
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Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.2 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |