+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-20272 | |||||||||
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Title | Structure of the K. lactis CBF3 core - Ndc10 D1D2 complex | |||||||||
Map data | K. lactis CBF3 core - Ndc10 D1D2 complex | |||||||||
Sample |
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Keywords | yeast centromere-binding complex / DNA BINDING PROTEIN | |||||||||
Function / homology | Function and homology information SCF-dependent proteasomal ubiquitin-dependent protein catabolic process / cullin family protein binding / mitotic cell cycle / DNA-binding transcription factor activity, RNA polymerase II-specific / protein ubiquitination / DNA binding / zinc ion binding / membrane / nucleus / cytoplasm Similarity search - Function | |||||||||
Biological species | Kluyveromyces lactis (yeast) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.3 Å | |||||||||
Authors | Lee PD / Wei H | |||||||||
Funding support | United States, 2 items
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Citation | Journal: J Mol Biol / Year: 2019 Title: Structure of the Centromere Binding Factor 3 Complex from Kluyveromyces lactis. Authors: Phong D Lee / Hui Wei / Dongyan Tan / Stephen C Harrison / Abstract: Kinetochores are the multiprotein complexes that link chromosomal centromeres to mitotic-spindle microtubules. Budding yeast centromeres comprise three sequential "centromere-determining elements", ...Kinetochores are the multiprotein complexes that link chromosomal centromeres to mitotic-spindle microtubules. Budding yeast centromeres comprise three sequential "centromere-determining elements", CDEI, II, and III. CDEI (8 bp) and CDEIII (∼25 bp) are conserved between Kluyveromyces lactis and Saccharomyces cerevisiae, but CDEII in the former is twice as long (160 bp) as CDEII in the latter (80 bp). The CBF3 complex recognizes CDEIII and is required for assembly of a centromeric nucleosome, which in turn recruits other kinetochore components. To understand differences in centromeric nucleosome assembly between K. lactis and S. cerevisiae, we determined the structure of a K. lactis CBF3 complex by electron cryomicroscopy at ∼4 Å resolution and compared it with published structures of S. cerevisiae CBF3. We show differences in the pose of Ndc10 and discuss potential models of the K. lactis centromeric nucleosome that account for the extended CDEII length. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_20272.map.gz | 5.6 MB | EMDB map data format | |
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Header (meta data) | emd-20272-v30.xml emd-20272.xml | 18.2 KB 18.2 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_20272_fsc.xml | 10.3 KB | Display | FSC data file |
Images | emd_20272.png | 61 KB | ||
Filedesc metadata | emd-20272.cif.gz | 6.2 KB | ||
Others | emd_20272_half_map_1.map.gz emd_20272_half_map_2.map.gz | 71.4 MB 71.2 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-20272 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-20272 | HTTPS FTP |
-Validation report
Summary document | emd_20272_validation.pdf.gz | 790.1 KB | Display | EMDB validaton report |
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Full document | emd_20272_full_validation.pdf.gz | 789.7 KB | Display | |
Data in XML | emd_20272_validation.xml.gz | 17.8 KB | Display | |
Data in CIF | emd_20272_validation.cif.gz | 23 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-20272 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-20272 | HTTPS FTP |
-Related structure data
Related structure data | 6p7xMC 6p7vC 6p7wC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_20272.map.gz / Format: CCP4 / Size: 91.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | K. lactis CBF3 core - Ndc10 D1D2 complex | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.0605 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Half map: K. lactis CBF3 core - Ndc10 D1D2 complex, half map 1
File | emd_20272_half_map_1.map | ||||||||||||
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Annotation | K. lactis CBF3 core - Ndc10 D1D2 complex, half map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: K. lactis CBF3 core - Ndc10 D1D2 complex, half map 2
File | emd_20272_half_map_2.map | ||||||||||||
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Annotation | K. lactis CBF3 core - Ndc10 D1D2 complex, half map 2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : K. lactis CBF3 core - Ndc10 D1D2 Complex
Entire | Name: K. lactis CBF3 core - Ndc10 D1D2 Complex |
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Components |
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-Supramolecule #1: K. lactis CBF3 core - Ndc10 D1D2 Complex
Supramolecule | Name: K. lactis CBF3 core - Ndc10 D1D2 Complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Kluyveromyces lactis (yeast) |
-Macromolecule #1: Ndc10
Macromolecule | Name: Ndc10 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Kluyveromyces lactis (yeast) |
Molecular weight | Theoretical: 47.053719 KDa |
Recombinant expression | Organism: Escherichia coli (E. coli) |
Sequence | String: MSKLDSLLKE LPTRTAHLYR SIWHKYTEWL KTMPDLTGAD LKLFLSQKYI VKYIASHDDI AKDPLPTCDA MIWFSRALDI ENNDVLVLQ QRLYGLVKLL EFDYSNVIAI LQKISINLWN PSTDSLQSKH FKTCQDKLKL LLDFQWKFNT NVSFEDRTTV S LKDLQCIL ...String: MSKLDSLLKE LPTRTAHLYR SIWHKYTEWL KTMPDLTGAD LKLFLSQKYI VKYIASHDDI AKDPLPTCDA MIWFSRALDI ENNDVLVLQ QRLYGLVKLL EFDYSNVIAI LQKISINLWN PSTDSLQSKH FKTCQDKLKL LLDFQWKFNT NVSFEDRTTV S LKDLQCIL DDENGKCGLA HSSKPNFVLV PNFQSPFTCP IFTMAVYYYL RFHGVKKYYK GDGYQILSQL EHIPIIRGKS LD QYPRELT LGNWYPTIFK YCQLPYTKKH WFQVNQEWPQ FPDFSDSSEN TSTLAESDSE NTIGIPDFYI EKMNRTKLQP CPQ VHVHLF PTDLPPDIQA VFDLLNSVLV TSLPLLYRVF PTHDIFLDPS LKTPQNIAFL TGTLPLDIES QEHLLAQLID KTGT VSEIG SG UniProtKB: KLLA0E03807p |
-Macromolecule #2: Ctf13
Macromolecule | Name: Ctf13 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Kluyveromyces lactis (yeast) |
Molecular weight | Theoretical: 46.159945 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: MFDTKLFLSL PIDIRYTVYF FLGDVVQNVR PPAKSDIFND ELIAYPNIRE FNQSLVDKYS KHIGVYDYIP NFIPNWCRDF DLLRHDIIL TDRLRVCLQY EEQWFSVQWI VVSGELEIGI FTTDEQFLQV SYTINEYCHL LSIAQQDLRL GINVSDINDV N ELCKEIQH ...String: MFDTKLFLSL PIDIRYTVYF FLGDVVQNVR PPAKSDIFND ELIAYPNIRE FNQSLVDKYS KHIGVYDYIP NFIPNWCRDF DLLRHDIIL TDRLRVCLQY EEQWFSVQWI VVSGELEIGI FTTDEQFLQV SYTINEYCHL LSIAQQDLRL GINVSDINDV N ELCKEIQH RWLFDTVSYI SFINCWDLDH ENVVSIIPCM ESFNNLHMLR IESKNMFNNL INTQGVRENP GKTIVYNVRQ NI FELELYT LRDLGYKSVV DLQKWEQLQC LSLSGCEFID LNNLILPQHC KMLILKEVKY IIWWDLSHLL KRIRPQWIIN GQV KKPTKK EEEEESEWYN LYLEVVQTYQ PLNFIELHNA KRVKGNLILP ARLVTESRIK ISNGTKVDSV LLI UniProtKB: KLLA0F13816p |
-Macromolecule #3: Skp1
Macromolecule | Name: Skp1 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Kluyveromyces lactis (yeast) |
Molecular weight | Theoretical: 21.081141 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: MSKENQNVVL VSVEGERFVV DRKIAERSLL LKNYLQDLNS GDLHDDNDAD DDEDDEEDGD DEIVMPVPNV RSSVLQKVIE WAVHHKDSN FPDEDDDDSR KAAPVDPWDR EFLKVDQEML YEIILAANYL NIKPLLDAGC KVVAEMIRGR TPEEIRRTFN I VNDFTPEE EAAIRRENEW AEDR UniProtKB: E3 ubiquitin ligase complex SCF subunit |
-Macromolecule #4: Cep3
Macromolecule | Name: Cep3 / type: protein_or_peptide / ID: 4 / Number of copies: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: Kluyveromyces lactis (yeast) |
Molecular weight | Theoretical: 74.165312 KDa |
Recombinant expression | Organism: Trichoplusia ni (cabbage looper) |
Sequence | String: MSKPKISLTK GKHPCTFCQA RKVKCDRSLP ACQNCIERNV TELCEYDDNG SRKRARLADD VNLYDKKLFN IWNQYERLWI HDTLGQCQQ GVYMGIAFPL DVSEYNNTKD FYGYECLFSK ESIFKILDHS LERLGWLYFG FFTDISELPY QMERYWNEYE S MNINLENE ...String: MSKPKISLTK GKHPCTFCQA RKVKCDRSLP ACQNCIERNV TELCEYDDNG SRKRARLADD VNLYDKKLFN IWNQYERLWI HDTLGQCQQ GVYMGIAFPL DVSEYNNTKD FYGYECLFSK ESIFKILDHS LERLGWLYFG FFTDISELPY QMERYWNEYE S MNINLENE EATTRQTTFK KSADQILWDL VLRSVIVMTI YYMPAKSILS LVDIDAIEKY PLDFSESNEG VDELKKKYEI FD YCLRHTL NKVLRTIFTL PPDVRTLQIF LILSNTNFLQ IYPSLGNNIL VHCIHLAKVL GIKDFKLKIN DSGSTRLQKL SMH NIWFRL STVDYMRSSP NKIIALHTDN SSALTRKTLF THCSIDSIDV YDVESNLEVL RWKITSLDRD LEVSEPSLKT LKAM KELLG LLDRKTSVSN DASFNTKFES FFLKLQCNFV MWKILRYEFM QYGVTNGLQK LCCPARRIIA LVANFLKEDY FEYTT HPFC VHILCVIAGF FSFYCIFHEA DEVRDLRNDA VGLLKLLFDP LRPVISCFFS NLSRLEELRH IWKSVEITDQ ANRLVH PVM YVLKTDIIKL KRNLEIISGS LKDANYQETF KDKLEIDINT PALSSDFLEV VREFNLSHPL DINGKMSRQN N UniProtKB: KLLA0D09977p |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 8 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 65.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |