+
Open data
-
Basic information
| Entry | ![]() | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Title | cryo-EM structure of Mis151-249-Cnp1 nucleosome complex | |||||||||
Map data | cryo-EM structure of Mis151-249-Cnp1 nucleosome complex | |||||||||
Sample |
| |||||||||
Keywords | Nucleosome dynamics / CENP-A / Cnp1 / kinetochore assembly / Chromosome Segregation / NUCLEAR PROTEIN/DNA / NUCLEAR PROTEIN-DNA complex | |||||||||
| Function / homology | Function and homology informationPKMTs methylate histone lysines / HDMs demethylate histones / CENP-A containing chromatin / : / Condensation of Prophase Chromosomes / HATs acetylate histones / : / : / Assembly of the ORC complex at the origin of replication / Oxidative Stress Induced Senescence ...PKMTs methylate histone lysines / HDMs demethylate histones / CENP-A containing chromatin / : / Condensation of Prophase Chromosomes / HATs acetylate histones / : / : / Assembly of the ORC complex at the origin of replication / Oxidative Stress Induced Senescence / Factors involved in megakaryocyte development and platelet production / RMTs methylate histone arginines / Ub-specific processing proteases / HDACs deacetylate histones / chromosome, centromeric core domain / SUMOylation of chromatin organization proteins / Estrogen-dependent gene expression / RNA Polymerase I Promoter Escape / Recruitment and ATM-mediated phosphorylation of repair and signaling proteins at DNA double strand breaks / Transcriptional regulation by small RNAs / Mis6-Sim4 complex / mating-type region heterochromatin / mitotic DNA damage checkpoint signaling / chromosome, subtelomeric region / CENP-A containing chromatin assembly / centromeric DNA binding / homologous chromosome segregation / rDNA heterochromatin / condensed chromosome, centromeric region / mitotic chromosome condensation / chromatin-protein adaptor activity / mitotic G2 DNA damage checkpoint signaling / chromosome, centromeric region / pericentric heterochromatin / CENP-A containing nucleosome / chromosome segregation / double-strand break repair via homologous recombination / kinetochore / structural constituent of chromatin / heterochromatin formation / nucleosome / double-strand break repair / nucleosome assembly / site of double-strand break / chromatin remodeling / protein heterodimerization activity / cell division / chromatin / DNA binding / nucleus Similarity search - Function | |||||||||
| Biological species | ![]() ![]() | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.83 Å | |||||||||
Authors | Xiong Y / Zang J | |||||||||
| Funding support | 1 items
| |||||||||
Citation | Journal: J Mol Cell Biol / Year: 2025Title: Cnp1 N-terminal dynamics regulate L1 loop recognition by Mis15 to orchestrate kinetochore assembly in Schizosaccharomyces pombe. Authors: Yujie Xiong / Yanze Jian / Yongliang Zhang / Min Zhang / Xuan Zhang / Kaiming Zhang / Chuanhai Fu / Tian Tian / Jianye Zang / ![]() Abstract: Centromeres are defined by the histone H3 variant CENP-A, which serve as the foundation for kinetochore assembly and ensure faithful chromosome segregation. CENP-A nucleosomes possess distinctive ...Centromeres are defined by the histone H3 variant CENP-A, which serve as the foundation for kinetochore assembly and ensure faithful chromosome segregation. CENP-A nucleosomes possess distinctive dynamic features, including flexible DNA ends at the entry/exit sites and a mobile N-terminal region, which are properties proposed to facilitate kinetochore assembly, yet the underlying molecular mechanisms remain elusive. Here, we present cryo-electron microscopy structures of Cnp1, the Schizosaccharomyces pombe (S. pombe) ortholog of CENP-A, alone and in complex with Mis15, the fission yeast ortholog of CENP-N. By integrating structural, biochemical, and molecular dynamics analyses, we demonstrate that the N-terminal region of Cnp1 regulates both DNA-end breathing and the conformational mobility of the L1 loop, a critical structural element for Mis15 recognition. Either enhanced dynamics caused by N-terminal deletion or reduced dynamics from targeted residue substitution disrupt Mis15 binding in vitro and impair its centromeric localization in vivo, thereby compromising the earliest steps of constitutive centromere-associated network assembly. Our findings establish the Cnp1 N-terminus as a dynamic allosteric modulator of chromatin architecture and reveal an L1 loop modulation mechanism that links nucleosome flexibility to kinetochore specification and chromosome segregation fidelity in fission yeast. | |||||||||
| History |
|
-
Structure visualization
| Supplemental images |
|---|
-
Downloads & links
-EMDB archive
| Map data | emd_63346.map.gz | 72.5 MB | EMDB map data format | |
|---|---|---|---|---|
| Header (meta data) | emd-63346-v30.xml emd-63346.xml | 25.1 KB 25.1 KB | Display Display | EMDB header |
| Images | emd_63346.png | 63.4 KB | ||
| Filedesc metadata | emd-63346.cif.gz | 6.5 KB | ||
| Others | emd_63346_additional_1.map.gz emd_63346_half_map_1.map.gz emd_63346_half_map_2.map.gz | 136.6 MB 134.3 MB 134.3 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-63346 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-63346 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9lrx ![]() 9lrvC ![]() 9lrwC M: atomic model generated by this map C: citing same article ( |
|---|---|
| Similar structure data | Similarity search - Function & homology F&H Search |
-
Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
|---|---|
| Related items in Molecule of the Month |
-
Map
| File | Download / File: emd_63346.map.gz / Format: CCP4 / Size: 144.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Annotation | cryo-EM structure of Mis151-249-Cnp1 nucleosome complex | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.07 Å | ||||||||||||||||||||||||||||||||||||
| Density |
| ||||||||||||||||||||||||||||||||||||
| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
|
-Supplemental data
-Additional map: cryo-EM structure of Mis151-249-Cnp1 nucleosome complex
| File | emd_63346_additional_1.map | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Annotation | cryo-EM structure of Mis151-249-Cnp1 nucleosome complex | ||||||||||||
| Projections & Slices |
| ||||||||||||
| Density Histograms |
-Half map: cryo-EM structure of Mis151-249-Cnp1 nucleosome complex
| File | emd_63346_half_map_1.map | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Annotation | cryo-EM structure of Mis151-249-Cnp1 nucleosome complex | ||||||||||||
| Projections & Slices |
| ||||||||||||
| Density Histograms |
-Half map: cryo-EM structure of Mis151-249-Cnp1 nucleosome complex
| File | emd_63346_half_map_2.map | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Annotation | cryo-EM structure of Mis151-249-Cnp1 nucleosome complex | ||||||||||||
| Projections & Slices |
| ||||||||||||
| Density Histograms |
-
Sample components
-Entire : cryo-EM structure of Mis151-249-Cnp1 nucleosome complex
| Entire | Name: cryo-EM structure of Mis151-249-Cnp1 nucleosome complex |
|---|---|
| Components |
|
-Supramolecule #1: cryo-EM structure of Mis151-249-Cnp1 nucleosome complex
| Supramolecule | Name: cryo-EM structure of Mis151-249-Cnp1 nucleosome complex type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
|---|---|
| Source (natural) | Organism: ![]() |
-Macromolecule #1: Inner kinetochore subunit mis15
| Macromolecule | Name: Inner kinetochore subunit mis15 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
|---|---|
| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 29.200105 KDa |
| Recombinant expression | Organism: in vitro transcription vector pT7-Fluc(deltai) (others) |
| Sequence | String: MRTELSSNIR IKHDKKIQKL LNRFPRDFLV KLCVEWIQKQ TYPPNAKDIN LEDMLDDEEW NPEAFYKNVP KSMLKRSIIH RMLVYDWPN GFYLGQIAQL EILALAHGFV SMRWTASKVH HSAEKTVLPN PLVFLELLKS ELESIFVYHT YISRHETLPI T FIRLVLWD ...String: MRTELSSNIR IKHDKKIQKL LNRFPRDFLV KLCVEWIQKQ TYPPNAKDIN LEDMLDDEEW NPEAFYKNVP KSMLKRSIIH RMLVYDWPN GFYLGQIAQL EILALAHGFV SMRWTASKVH HSAEKTVLPN PLVFLELLKS ELESIFVYHT YISRHETLPI T FIRLVLWD SKRPTALHTY PSSKQIFYLG LMDDSDVLLH NIFLRNDVCH SLFLQCLSRL LYRLKAGSAL RPIDLVSKNL TT FCTNVGV NK UniProtKB: Inner kinetochore subunit mis15 |
-Macromolecule #2: Histone H3-like centromeric protein cnp1
| Macromolecule | Name: Histone H3-like centromeric protein cnp1 / type: protein_or_peptide / ID: 2 / Number of copies: 2 / Enantiomer: LEVO |
|---|---|
| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 13.908231 KDa |
| Recombinant expression | Organism: in vitro transcription vector pT7-Fluc(deltai) (others) |
| Sequence | String: MAKKSLMAEP GDPIPRPRKK RYRPGTTALR EIRKYQRSTD LLIQRLPFSR IVREISSEFV ANFSTDVGLR WQSTALQCLQ EAAEAFLVH LFEDTNLCAI HAKRVTIMQR DMQLARRIRG A UniProtKB: Histone H3-like centromeric protein cnp1 |
-Macromolecule #3: Histone H4
| Macromolecule | Name: Histone H4 / type: protein_or_peptide / ID: 3 / Number of copies: 2 / Enantiomer: LEVO |
|---|---|
| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 8.952407 KDa |
| Recombinant expression | Organism: in vitro transcription vector pT7-Fluc(deltai) (others) |
| Sequence | String: DNIQGITKPA IRRLARRGGV KRISALVYEE TRAVLKLFLE NVIRDAVTYT EHAKRKTVTS LDVVYSLKRQ GRTIYGFGG UniProtKB: Histone H4 |
-Macromolecule #4: Histone H2A-beta
| Macromolecule | Name: Histone H2A-beta / type: protein_or_peptide / ID: 4 / Number of copies: 2 / Enantiomer: LEVO |
|---|---|
| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 11.295081 KDa |
| Recombinant expression | Organism: in vitro transcription vector pT7-Fluc(deltai) (others) |
| Sequence | String: SAQSRSAKAG LAFPVGRVHR LLRKGNYAQR VGAGAPVYLA AVLEYLAAEI LELAGNAARD NKKTRIIPRH LQLAIRNDEE LNKLLGHVT IAQGGVVPNI NAHLLP UniProtKB: Histone H2A-beta |
-Macromolecule #5: Histone H2B-alpha
| Macromolecule | Name: Histone H2B-alpha / type: protein_or_peptide / ID: 5 / Number of copies: 2 / Enantiomer: LEVO |
|---|---|
| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 10.540012 KDa |
| Recombinant expression | Organism: in vitro transcription vector pT7-Fluc(deltai) (others) |
| Sequence | String: KNRKETYSSY IYKVLKQVHP DTGISNQAMR ILNSFVNDIF ERIATEASKL AAYNKKSTIS SREIQTAVRL ILPGELAKHA VTEGTKSVT KYSSS UniProtKB: Histone H2B-alpha |
-Macromolecule #6: DNA (144-MER)
| Macromolecule | Name: DNA (144-MER) / type: dna / ID: 6 / Number of copies: 1 / Classification: DNA |
|---|---|
| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 44.168133 KDa |
| Sequence | String: (DT)(DC)(DG)(DA)(DG)(DA)(DA)(DT)(DC)(DC) (DC)(DG)(DG)(DT)(DG)(DC)(DC)(DG)(DA)(DG) (DG)(DC)(DC)(DG)(DC)(DT)(DC)(DA)(DA) (DT)(DT)(DG)(DG)(DT)(DC)(DG)(DT)(DA)(DG) (DA) (DC)(DA)(DG)(DC)(DT)(DC) ...String: (DT)(DC)(DG)(DA)(DG)(DA)(DA)(DT)(DC)(DC) (DC)(DG)(DG)(DT)(DG)(DC)(DC)(DG)(DA)(DG) (DG)(DC)(DC)(DG)(DC)(DT)(DC)(DA)(DA) (DT)(DT)(DG)(DG)(DT)(DC)(DG)(DT)(DA)(DG) (DA) (DC)(DA)(DG)(DC)(DT)(DC)(DT)(DA) (DG)(DC)(DA)(DC)(DC)(DG)(DC)(DT)(DT)(DA) (DA)(DA) (DC)(DG)(DC)(DA)(DC)(DG)(DT) (DA)(DC)(DG)(DC)(DG)(DC)(DT)(DG)(DT)(DC) (DC)(DC)(DC) (DC)(DG)(DC)(DG)(DT)(DT) (DT)(DT)(DA)(DA)(DC)(DC)(DG)(DC)(DC)(DA) (DA)(DG)(DG)(DG) (DG)(DA)(DT)(DT)(DA) (DC)(DT)(DC)(DC)(DC)(DT)(DA)(DG)(DT)(DC) (DT)(DC)(DC)(DA)(DG) (DG)(DC)(DA)(DC) (DG)(DT)(DG)(DT)(DC)(DA)(DC)(DA)(DT)(DA) (DT)(DA)(DT)(DA)(DC)(DA) (DT)(DC)(DC) (DG) |
-Macromolecule #7: DNA (144-MER)
| Macromolecule | Name: DNA (144-MER) / type: dna / ID: 7 / Number of copies: 1 / Classification: DNA |
|---|---|
| Source (natural) | Organism: ![]() |
| Molecular weight | Theoretical: 44.728477 KDa |
| Sequence | String: (DC)(DG)(DG)(DA)(DT)(DG)(DT)(DA)(DT)(DA) (DT)(DA)(DT)(DG)(DT)(DG)(DA)(DC)(DA)(DC) (DG)(DT)(DG)(DC)(DC)(DT)(DG)(DG)(DA) (DG)(DA)(DC)(DT)(DA)(DG)(DG)(DG)(DA)(DG) (DT) (DA)(DA)(DT)(DC)(DC)(DC) ...String: (DC)(DG)(DG)(DA)(DT)(DG)(DT)(DA)(DT)(DA) (DT)(DA)(DT)(DG)(DT)(DG)(DA)(DC)(DA)(DC) (DG)(DT)(DG)(DC)(DC)(DT)(DG)(DG)(DA) (DG)(DA)(DC)(DT)(DA)(DG)(DG)(DG)(DA)(DG) (DT) (DA)(DA)(DT)(DC)(DC)(DC)(DC)(DT) (DT)(DG)(DG)(DC)(DG)(DG)(DT)(DT)(DA)(DA) (DA)(DA) (DC)(DG)(DC)(DG)(DG)(DG)(DG) (DG)(DA)(DC)(DA)(DG)(DC)(DG)(DC)(DG)(DT) (DA)(DC)(DG) (DT)(DG)(DC)(DG)(DT)(DT) (DT)(DA)(DA)(DG)(DC)(DG)(DG)(DT)(DG)(DC) (DT)(DA)(DG)(DA) (DG)(DC)(DT)(DG)(DT) (DC)(DT)(DA)(DC)(DG)(DA)(DC)(DC)(DA)(DA) (DT)(DT)(DG)(DA)(DG) (DC)(DG)(DG)(DC) (DC)(DT)(DC)(DG)(DG)(DC)(DA)(DC)(DC)(DG) (DG)(DG)(DA)(DT)(DT)(DC) (DT)(DC)(DG) (DA) |
-Experimental details
-Structure determination
| Method | cryo EM |
|---|---|
Processing | single particle reconstruction |
| Aggregation state | particle |
-
Sample preparation
| Concentration | 0.2 mg/mL |
|---|---|
| Buffer | pH: 7.5 |
| Vitrification | Cryogen name: ETHANE |
-
Electron microscopy
| Microscope | FEI POLARA 300 |
|---|---|
| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 55.0 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: 2.0 µm / Nominal defocus min: 1.0 µm |
| Experimental equipment | ![]() Model: Tecnai Polara / Image courtesy: FEI Company |
Movie
Controller
About Yorodumi




Keywords
Authors
Citation







Z (Sec.)
Y (Row.)
X (Col.)












































Processing
FIELD EMISSION GUN
