+ Open data
Open data
- Basic information
Basic information
| Entry | Database: EMDB / ID: EMD-23823 | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Title | Closed linker DNA nucleosome reconstituted with GUB DNA | |||||||||
|  Map data | ||||||||||
|  Sample | 
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| Function / homology |  Function and homology information structural constituent of chromatin / nucleosome / heterochromatin formation / nucleosome assembly / protein heterodimerization activity / DNA binding / nucleoplasm / nucleus Similarity search - Function | |||||||||
| Biological species |  Xenopus laevis (African clawed frog) / synthetic construct (others) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.77 Å | |||||||||
|  Authors | Arimura Y / Funabiki H | |||||||||
| Funding support |  United States,  Japan, 2 items 
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|  Citation |  Journal: Mol Cell / Year: 2021 Title: Structural features of nucleosomes in interphase and metaphase chromosomes. Authors: Yasuhiro Arimura / Rochelle M Shih / Ruby Froom / Hironori Funabiki /  Abstract: Structural heterogeneity of nucleosomes in functional chromosomes is unknown. Here, we devise the template-, reference- and selection-free (TRSF) cryo-EM pipeline to simultaneously reconstruct cryo- ...Structural heterogeneity of nucleosomes in functional chromosomes is unknown. Here, we devise the template-, reference- and selection-free (TRSF) cryo-EM pipeline to simultaneously reconstruct cryo-EM structures of protein complexes from interphase or metaphase chromosomes. The reconstructed interphase and metaphase nucleosome structures are on average indistinguishable from canonical nucleosome structures, despite DNA sequence heterogeneity, cell-cycle-specific posttranslational modifications, and interacting proteins. Nucleosome structures determined by a decoy-classifying method and structure variability analyses reveal the nucleosome structural variations in linker DNA, histone tails, and nucleosome core particle configurations, suggesting that the opening of linker DNA, which is correlated with H2A C-terminal tail positioning, is suppressed in chromosomes. High-resolution (3.4-3.5 Å) nucleosome structures indicate DNA-sequence-independent stabilization of superhelical locations ±0-1 and ±3.5-4.5. The linker histone H1.8 preferentially binds to metaphase chromatin, from which chromatosome cryo-EM structures with H1.8 at the on-dyad position are reconstituted. This study presents the structural characteristics of nucleosomes in chromosomes. | |||||||||
| History | 
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- Structure visualization
Structure visualization
| Movie | 
 
  Movie viewer | 
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| Structure viewer | EM map:  SurfView  Molmil  Jmol/JSmol | 
| Supplemental images | 
- Downloads & links
Downloads & links
-EMDB archive
| Map data |  emd_23823.map.gz | 28.5 MB |  EMDB map data format | |
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| Header (meta data) |  emd-23823-v30.xml  emd-23823.xml | 19.3 KB 19.3 KB | Display Display |  EMDB header | 
| FSC (resolution estimation) |  emd_23823_fsc.xml | 7.2 KB | Display |  FSC data file | 
| Images |  emd_23823.png | 98.6 KB | ||
| Others |  emd_23823_half_map_1.map.gz  emd_23823_half_map_2.map.gz | 23.3 MB 23.3 MB | ||
| Archive directory |  http://ftp.pdbj.org/pub/emdb/structures/EMD-23823  ftp://ftp.pdbj.org/pub/emdb/structures/EMD-23823 | HTTPS FTP | 
-Validation report
| Summary document |  emd_23823_validation.pdf.gz | 527.4 KB | Display |  EMDB validaton report | 
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| Full document |  emd_23823_full_validation.pdf.gz | 527.1 KB | Display | |
| Data in XML |  emd_23823_validation.xml.gz | 14.1 KB | Display | |
| Data in CIF |  emd_23823_validation.cif.gz | 17.9 KB | Display | |
| Arichive directory |  https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23823  ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23823 | HTTPS FTP | 
-Related structure data
| Related structure data |  7kbdC  7kbeC  7kbfC C: citing same article ( | 
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| Similar structure data | |
| EM raw data |  EMPIAR-10749 (Title: Closed linker DNA nucleosome reconstituted with GUB DNA Data size: 624.4 Data #1: GUB DNA nucleosome with "closed" linker DNA [micrographs - multiframe]) | 
- Links
Links
| EMDB pages |  EMDB (EBI/PDBe) /  EMDataResource | 
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| Related items in Molecule of the Month | 
- Map
Map
| File |  Download / File: emd_23823.map.gz / Format: CCP4 / Size: 30.5 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: 1.47 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density | 
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML: 
 CCP4 map header: 
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-Supplemental data
-Half map: #1
| File | emd_23823_half_map_1.map | ||||||||||||
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| Projections & Slices | 
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| Density Histograms | 
-Half map: #2
| File | emd_23823_half_map_2.map | ||||||||||||
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| Projections & Slices | 
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| Density Histograms | 
- Sample components
Sample components
-Entire : Closed linker DNA nucleosome reconstituted with GUB DNA
| Entire | Name: Closed linker DNA nucleosome reconstituted with GUB DNA | 
|---|---|
| Components | 
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-Supramolecule #1: Closed linker DNA nucleosome reconstituted with GUB DNA
| Supramolecule | Name: Closed linker DNA nucleosome reconstituted with GUB DNA type: complex / ID: 1 / Parent: 0 / Macromolecule list: all | 
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| Source (natural) | Organism:  Xenopus laevis (African clawed frog) | 
-Macromolecule #1: H3.2
| Macromolecule | Name: H3.2 / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO | 
|---|---|
| Source (natural) | Organism:  Xenopus laevis (African clawed frog) | 
| Recombinant expression | Organism:   Escherichia coli (E. coli) | 
| Sequence | String: ARTKQTARKS TGGKAPRKQL ATKAARKSAP ATGGVKKPHR YRPGTVALRE IRRYQKSTEL LIRKLPFQRL VREIAQDFKT DLRFQSSAVM ALQEASEAYL VALFEDTNLC AIHAKRVTIM PKDIQLARRI RGERA | 
-Macromolecule #2: H4
| Macromolecule | Name: H4 / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO | 
|---|---|
| Source (natural) | Organism:  Xenopus laevis (African clawed frog) | 
| Recombinant expression | Organism:   Escherichia coli (E. coli) | 
| Sequence | String: SGRGKGGKGL GKGGAKRHRK VLRDNIQGIT KPAIRRLARR GGVKRISGLI YEETRGVLKV FLENVIRDAV TYTEHAKRKT VTAMDVVYAL KRQGRTLYGF GG | 
-Macromolecule #3: H2A
| Macromolecule | Name: H2A / type: protein_or_peptide / ID: 3 / Enantiomer: LEVO | 
|---|---|
| Source (natural) | Organism:  Xenopus laevis (African clawed frog) | 
| Recombinant expression | Organism:   Escherichia coli (E. coli) | 
| Sequence | String: SGRGKQGGKT RAKAKTRSSR AGLQFPVGRV HRLLRKGNYA ERVGAGAPVY LAAVLEYLTA EILELAGNAA RDNKKTRIIP RHLQLAVRND EELNKLLGRV TIAQGGVLPN IQSVLLPKKT ESSKSAKSK | 
-Macromolecule #4: H2B
| Macromolecule | Name: H2B / type: protein_or_peptide / ID: 4 / Enantiomer: LEVO | 
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| Source (natural) | Organism:  Xenopus laevis (African clawed frog) | 
| Recombinant expression | Organism:   Escherichia coli (E. coli) | 
| Sequence | String: MPEPAKSAPA PKKGSKKAVT KTQKKDGKKR RKTRKESYAI YVYKVLKQVH PDTGISSKAM SIMNSFVNDV FERIAGEASR LAHYNKRSTI TSREIQTAVR LLLPGELAKH AVSEGTKAVT KYTSAK | 
-Macromolecule #5: GUB DNA
| Macromolecule | Name: GUB DNA / type: dna / ID: 5 / Classification: DNA | 
|---|---|
| Source (natural) | Organism: synthetic construct (others) | 
| Sequence | String: ATCCCTCTAG ACGGAGGACA GTCCTCCGGT TACCTTCGAA CCACGTGGCC GTCTAGATGC TGACTCATTG TCGACACGCG TAGATCTGCT AGCATCGATC CATGGACTAG TCTCGAGTTT AAAGATATCC AGCTGCCCGG GAGGCCTTCG CGAAATATTG GTACCCCATG GAAGAT | 
-Macromolecule #6: GUB DNA_2
| Macromolecule | Name: GUB DNA_2 / type: dna / ID: 6 / Classification: DNA | 
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| Source (natural) | Organism: synthetic construct (others) | 
| Sequence | String: ATCTTCCATG GGGTACCAAT ATTTCGCGAA GGCCTCCCGG GCAGCTGGAT ATCTTTAAAC TCGAGACTAG TCCATGGATC GATGCTAGCA GATCTACGCG TGTCGACAAT GAGTCAGCAT CTAGACGGCC ACGTGGTTCG AAGGTAACCG GAGGACTGTC CTCCGTCTAG AGGGAT | 
-Experimental details
-Structure determination
| Method | cryo EM | 
|---|---|
|  Processing | single particle reconstruction | 
| Aggregation state | particle | 
- Sample preparation
Sample preparation
| Buffer | pH: 7.4 | 
|---|---|
| Grid | Model: Quantifoil R1.2/1.3 / Material: GOLD | 
| Vitrification | Cryogen name: ETHANE | 
| Details | Closed linker DNA nucleosome reconstituted with GUB DNA | 
- Electron microscopy
Electron microscopy
| Microscope | FEI TALOS ARCTICA | 
|---|---|
| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Average electron dose: 35.5 e/Å2 | 
| Electron beam | Acceleration voltage: 200 kV / Electron source:  FIELD EMISSION GUN | 
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD | 
| Experimental equipment |  Model: Talos Arctica / Image courtesy: FEI Company | 
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