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Yorodumi- EMDB-33313: RNA polymerase II elongation complex containing Spt4/5, Elf1, Spt... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-33313 | ||||||||||||||||||||||||
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Title | RNA polymerase II elongation complex containing Spt4/5, Elf1, Spt6, Spn1 and Paf1C | ||||||||||||||||||||||||
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Keywords | chromatin / nucleosome / TRANSCRIPTION | ||||||||||||||||||||||||
Function / homology | Function and homology information RNA polymerase II C-terminal domain phosphoserine binding / nuclear DNA-directed RNA polymerase complex / regulation of septum digestion after cytokinesis / Cdc73/Paf1 complex / siRNA-mediated pericentric heterochromatin formation / DSIF complex / regulation of transcription elongation by RNA polymerase II / nucleosome organization / RPB4-RPB7 complex / transcription elongation factor activity ...RNA polymerase II C-terminal domain phosphoserine binding / nuclear DNA-directed RNA polymerase complex / regulation of septum digestion after cytokinesis / Cdc73/Paf1 complex / siRNA-mediated pericentric heterochromatin formation / DSIF complex / regulation of transcription elongation by RNA polymerase II / nucleosome organization / RPB4-RPB7 complex / transcription elongation factor activity / poly(A)+ mRNA export from nucleus / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / transcription elongation-coupled chromatin remodeling / termination of RNA polymerase II transcription / termination of RNA polymerase III transcription / transcription initiation at RNA polymerase III promoter / termination of RNA polymerase I transcription / RNA polymerase III activity / RNA polymerase II complex binding / transcription initiation at RNA polymerase I promoter / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / RNA polymerase II activity / chromosome, centromeric region / transcription elongation by RNA polymerase I / nucleosome binding / tRNA transcription by RNA polymerase III / RNA polymerase I complex / RNA polymerase I activity / RNA polymerase III complex / positive regulation of translational initiation / translation elongation factor activity / RNA polymerase II, core complex / pericentric heterochromatin / translation initiation factor binding / transcription elongation factor complex / regulation of DNA-templated transcription elongation / transcription initiation at RNA polymerase II promoter / positive regulation of transcription elongation by RNA polymerase II / transcription elongation by RNA polymerase II / P-body / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / single-stranded DNA binding / histone binding / nucleic acid binding / single-stranded RNA binding / protein dimerization activity / RNA-dependent RNA polymerase activity / mRNA binding / nucleotide binding / DNA-templated transcription / chromatin binding / regulation of DNA-templated transcription / regulation of transcription by RNA polymerase II / nucleolus / DNA binding / RNA binding / zinc ion binding / nucleoplasm / nucleus / metal ion binding Similarity search - Function | ||||||||||||||||||||||||
Biological species | Komagataella phaffii (fungus) / synthetic construct (others) | ||||||||||||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | ||||||||||||||||||||||||
Authors | Ehara H / Kujirai T / Shirouzu M / Kurumizaka H / Sekine S | ||||||||||||||||||||||||
Funding support | Japan, 7 items
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Citation | Journal: Science / Year: 2022 Title: Structural basis of nucleosome disassembly and reassembly by RNAPII elongation complex with FACT. Authors: Haruhiko Ehara / Tomoya Kujirai / Mikako Shirouzu / Hitoshi Kurumizaka / Shun-Ichi Sekine / Abstract: During gene transcription, RNA polymerase II (RNAPII) traverses nucleosomes in chromatin, but the mechanism has remained elusive. Using cryo-electron microscopy, we obtained structures of the RNAPII ...During gene transcription, RNA polymerase II (RNAPII) traverses nucleosomes in chromatin, but the mechanism has remained elusive. Using cryo-electron microscopy, we obtained structures of the RNAPII elongation complex (EC) passing through a nucleosome in the presence of the transcription elongation factors Spt6, Spn1, Elf1, Spt4/5, and Paf1C and the histone chaperone FACT (facilitates chromatin transcription). The structures show snapshots of EC progression on DNA mediating downstream nucleosome disassembly, followed by its reassembly upstream of the EC, which is facilitated by FACT. FACT dynamically adapts to successively occurring subnucleosome intermediates, forming an interface with the EC. Spt6, Spt4/5, and Paf1C form a "cradle" at the EC DNA-exit site and support the upstream nucleosome reassembly. These structures explain the mechanism by which the EC traverses nucleosomes while maintaining the chromatin structure and epigenetic information. | ||||||||||||||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_33313.map.gz | 40.6 MB | EMDB map data format | |
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Header (meta data) | emd-33313-v30.xml emd-33313.xml | 64.4 KB 64.4 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_33313_fsc.xml | 8.5 KB | Display | FSC data file |
Images | emd_33313.png | 130.8 KB | ||
Filedesc metadata | emd-33313.cif.gz | 14.5 KB | ||
Others | emd_33313_additional_1.map.gz emd_33313_additional_2.map.gz emd_33313_additional_3.map.gz emd_33313_additional_4.map.gz emd_33313_additional_5.map.gz emd_33313_additional_6.map.gz emd_33313_additional_7.map.gz emd_33313_half_map_1.map.gz emd_33313_half_map_2.map.gz | 40.4 MB 23.3 MB 40.6 MB 40.7 MB 40.6 MB 23.3 MB 40.7 MB 40.7 MB 40.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-33313 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-33313 | HTTPS FTP |
-Validation report
Summary document | emd_33313_validation.pdf.gz | 819.6 KB | Display | EMDB validaton report |
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Full document | emd_33313_full_validation.pdf.gz | 819.2 KB | Display | |
Data in XML | emd_33313_validation.xml.gz | 14 KB | Display | |
Data in CIF | emd_33313_validation.cif.gz | 20.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-33313 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-33313 | HTTPS FTP |
-Related structure data
Related structure data | 7xn7MC 7xseC 7xsxC 7xszC 7xt7C 7xtdC 7xtiC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_33313.map.gz / Format: CCP4 / Size: 52.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.484 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: Map3
File | emd_33313_additional_1.map | ||||||||||||
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Annotation | Map3 | ||||||||||||
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-Additional map: Map2
File | emd_33313_additional_2.map | ||||||||||||
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Annotation | Map2 | ||||||||||||
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-Additional map: Map1
File | emd_33313_additional_3.map | ||||||||||||
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Annotation | Map1 | ||||||||||||
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-Additional map: Map4
File | emd_33313_additional_4.map | ||||||||||||
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Annotation | Map4 | ||||||||||||
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-Additional map: Map5
File | emd_33313_additional_5.map | ||||||||||||
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Annotation | Map5 | ||||||||||||
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-Additional map: Map6
File | emd_33313_additional_6.map | ||||||||||||
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Annotation | Map6 | ||||||||||||
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-Additional map: Map7
File | emd_33313_additional_7.map | ||||||||||||
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Annotation | Map7 | ||||||||||||
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-Half map: #2
File | emd_33313_half_map_1.map | ||||||||||||
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-Half map: #1
File | emd_33313_half_map_2.map | ||||||||||||
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-Sample components
+Entire : RNA polymerase II elongation complex containing Spt4/5, Elf1, Spt...
+Supramolecule #1: RNA polymerase II elongation complex containing Spt4/5, Elf1, Spt...
+Macromolecule #1: DNA-directed RNA polymerase subunit
+Macromolecule #2: DNA-directed RNA polymerase subunit beta
+Macromolecule #3: RNA polymerase II third largest subunit B44, part of central core
+Macromolecule #4: RNA polymerase II subunit B32
+Macromolecule #5: DNA-directed RNA polymerases I, II, and III subunit RPABC1
+Macromolecule #6: RNA polymerase subunit ABC23, common to RNA polymerases I, II, and III
+Macromolecule #7: RNA polymerase II subunit
+Macromolecule #8: DNA-directed RNA polymerases I, II, and III subunit RPABC3
+Macromolecule #9: DNA-directed RNA polymerase subunit
+Macromolecule #10: RNA polymerase subunit ABC10-beta, common to RNA polymerases I, I...
+Macromolecule #11: RNA polymerase II subunit B12.5
+Macromolecule #12: RNA polymerase subunit ABC10-alpha
+Macromolecule #13: Transcription elongation factor 1 homolog
+Macromolecule #17: Transcription elongation factor SPT4
+Macromolecule #18: Chromatin elongation factor SPT5
+Macromolecule #19: Transcription elongation factor Spt6
+Macromolecule #20: Protein that interacts with Spt6p and copurifies with Spt5p and R...
+Macromolecule #21: Component of the Paf1p complex
+Macromolecule #22: RNAPII-associated chromatin remodeling Paf1 complex subunit
+Macromolecule #23: Leo1
+Macromolecule #24: RNAP II-associated protein
+Macromolecule #25: Constituent of Paf1 complex with RNA polymerase II, Paf1p, Hpr1p,...
+Macromolecule #14: DNA (198-mer)
+Macromolecule #16: DNA (198-mer)
+Macromolecule #15: RNA (5'-R(P*AP*AP*GP*CP*CP*UP*GP*GP*UP*GP*UP*CP*UP*UP*GP*GP*GP*UP...
+Macromolecule #26: ZINC ION
+Macromolecule #27: MAGNESIUM ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 51.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.0 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |