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Yorodumi- EMDB-33424: RNA polymerase II elongation complex transcribing a nucleosome (EC42) -
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Basic information
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| Title | RNA polymerase II elongation complex transcribing a nucleosome (EC42) | ||||||||||||||||||||||||
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Keywords | chromatin / nucleosome / TRANSCRIPTION | ||||||||||||||||||||||||
| Function / homology | Function and homology informationsnoRNA transcription by RNA polymerase II / regulation of transcription initiation by RNA polymerase II / negative regulation of transcription elongation by RNA polymerase I / sno(s)RNA 3'-end processing / positive regulation of transcription elongation by RNA polymerase I / mating-type region heterochromatin / transcription antitermination factor activity, DNA binding / regulation of septum digestion after cytokinesis / RNA polymerase II C-terminal domain phosphoserine binding / regulation of transcription-coupled nucleotide-excision repair ...snoRNA transcription by RNA polymerase II / regulation of transcription initiation by RNA polymerase II / negative regulation of transcription elongation by RNA polymerase I / sno(s)RNA 3'-end processing / positive regulation of transcription elongation by RNA polymerase I / mating-type region heterochromatin / transcription antitermination factor activity, DNA binding / regulation of septum digestion after cytokinesis / RNA polymerase II C-terminal domain phosphoserine binding / regulation of transcription-coupled nucleotide-excision repair / triplex DNA binding / Cdc73/Paf1 complex / co-transcriptional lncRNA 3' end processing, cleavage and polyadenylation pathway / siRNA-mediated pericentric heterochromatin formation / regulation of mRNA 3'-end processing / RNA polymerase I core binding / DSIF complex / regulation of rRNA processing / intracellular mRNA localization / RNA polymerase I general transcription initiation factor binding / rDNA binding / rDNA heterochromatin / negative regulation of DNA recombination / transcription elongation factor complex / intracellular phosphate ion homeostasis / negative regulation of mitophagy / transcription elongation-coupled chromatin remodeling / snRNP binding / mRNA 3'-end processing / U4 snRNA binding / transcription elongation factor activity / chromatin-protein adaptor activity / cellular response to stress / poly(A)+ mRNA export from nucleus / kinetochore assembly / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / positive regulation of nuclear-transcribed mRNA poly(A) tail shortening / spliceosomal complex assembly / RNA polymerase II complex binding / termination of RNA polymerase II transcription / U5 snRNA binding / 7-methylguanosine mRNA capping / termination of RNA polymerase I transcription / maintenance of transcriptional fidelity during transcription elongation by RNA polymerase II / transcription initiation at RNA polymerase I promoter / positive regulation of translational initiation / mitotic metaphase chromosome alignment / U2 snRNA binding / nuclear-transcribed mRNA catabolic process / U6 snRNA binding / negative regulation of tumor necrosis factor-mediated signaling pathway / translation elongation factor activity / U1 snRNA binding / pericentric heterochromatin / RNA polymerase II core promoter sequence-specific DNA binding / termination of RNA polymerase III transcription / transcription initiation at RNA polymerase III promoter / RNA polymerase I complex / nucleosome binding / RNA polymerase III complex / negative regulation of megakaryocyte differentiation / RNA polymerase II, core complex / transcription elongation by RNA polymerase I / tRNA transcription by RNA polymerase III / protein localization to CENP-A containing chromatin / transcription by RNA polymerase I / Replacement of protamines by nucleosomes in the male pronucleus / Packaging Of Telomere Ends / transcription-coupled nucleotide-excision repair / translation initiation factor binding / Recognition and association of DNA glycosylase with site containing an affected purine / Cleavage of the damaged purine / positive regulation of autophagy / negative regulation of autophagy / telomere organization / Deposition of new CENPA-containing nucleosomes at the centromere / Recognition and association of DNA glycosylase with site containing an affected pyrimidine / Cleavage of the damaged pyrimidine / translesion synthesis / transcription initiation-coupled chromatin remodeling / RNA Polymerase I Promoter Opening / Inhibition of DNA recombination at telomere / Assembly of the ORC complex at the origin of replication / SUMOylation of chromatin organization proteins / Regulation of endogenous retroelements by the Human Silencing Hub (HUSH) complex / Meiotic synapsis / DNA methylation / Condensation of Prophase Chromosomes / Chromatin modifications during the maternal to zygotic transition (MZT) / SIRT1 negatively regulates rRNA expression / HCMV Late Events / ERCC6 (CSB) and EHMT2 (G9a) positively regulate rRNA expression / PRC2 methylates histones and DNA / Regulation of endogenous retroelements by KRAB-ZFP proteins / G1/S transition of mitotic cell cycle / Defective pyroptosis / HDACs deacetylate histones / positive regulation of transcription elongation by RNA polymerase II / Transcriptional regulation by small RNAs / lipopolysaccharide binding Similarity search - Function | ||||||||||||||||||||||||
| Biological species | Komagataella phaffii (fungus) / Homo sapiens (human) / synthetic construct (others) | ||||||||||||||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.6 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: 2022Title: 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. | ||||||||||||||||||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_33424.map.gz | 40.8 MB | EMDB map data format | |
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| Header (meta data) | emd-33424-v30.xml emd-33424.xml | 63.5 KB 63.5 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_33424_fsc.xml | 8.5 KB | Display | FSC data file |
| Images | emd_33424.png | 109.8 KB | ||
| Filedesc metadata | emd-33424.cif.gz | 15.2 KB | ||
| Others | emd_33424_additional_1.map.gz emd_33424_additional_2.map.gz emd_33424_half_map_1.map.gz emd_33424_half_map_2.map.gz | 40.8 MB 40.4 MB 40.9 MB 40.9 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-33424 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-33424 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 7xseMC ![]() 7xn7C ![]() 7xsxC ![]() 7xszC ![]() 7xt7C ![]() 7xtdC ![]() 7xtiC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_33424.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: EC reconstruction
| File | emd_33424_additional_1.map | ||||||||||||
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| Annotation | EC reconstruction | ||||||||||||
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| Density Histograms |
-Additional map: nucleosome reconstruction
| File | emd_33424_additional_2.map | ||||||||||||
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| Annotation | nucleosome reconstruction | ||||||||||||
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-Half map: #2
| File | emd_33424_half_map_1.map | ||||||||||||
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| Density Histograms |
-Half map: #1
| File | emd_33424_half_map_2.map | ||||||||||||
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| Density Histograms |
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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...
+Supramolecule #2: RNA polymerase II
+Supramolecule #3: Elf1,Spt4/5,Spn1 and Paf1C
+Supramolecule #4: DNA,RNA
+Supramolecule #5: histone
+Supramolecule #6: Spt6
+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: Transcription elongation factor SPT5
+Macromolecule #19: Histone H3.3
+Macromolecule #20: Histone H4
+Macromolecule #21: Histone H2A type 1-B/E
+Macromolecule #22: Histone H2B type 1-J
+Macromolecule #23: Transcription elongation factor Spt6
+Macromolecule #24: Protein that interacts with Spt6p and copurifies with Spt5p and R...
+Macromolecule #25: Component of the Paf1p complex
+Macromolecule #26: RNAPII-associated chromatin remodeling Paf1 complex subunit
+Macromolecule #27: Leo1
+Macromolecule #28: RNAP II-associated protein
+Macromolecule #29: 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*UP*GP*CP*CP*UP*GP*GP*UP*GP*UP*CP*UP*UP*GP*GP*GP*UP*GP...
+Macromolecule #30: ZINC ION
+Macromolecule #31: MAGNESIUM ION
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 7.5 |
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| Vitrification | Cryogen name: ETHANE |
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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 |
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About Yorodumi



Keywords
Komagataella phaffii (fungus)
Homo sapiens (human)
Authors
Japan, 7 items
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Processing
FIELD EMISSION GUN

