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- EMDB-17224: Cryo-EM structure of CBF1-CCAN bound topologically to centromeric DNA -

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Basic information

Entry
Database: EMDB / ID: EMD-17224
TitleCryo-EM structure of CBF1-CCAN bound topologically to centromeric DNA
Map data
Sample
  • Complex: A complex of CBF1-CCAN bound to centromeric C0N3 DNA
    • Protein or peptide: x 14 types
    • DNA: x 2 types
Keywordskinetochore / point centromere / CENP-A nucleosome / topological entrapment / centromeric DNA / CELL CYCLE
Function / homology
Function and homology information


Cbf1-Met4-Met28 complex / regulation of sulfur metabolic process / negative regulation of kinetochore assembly / negative regulation of meiotic DNA double-strand break formation involved in reciprocal meiotic recombination / COMA complex / maintenance of meiotic sister chromatid cohesion / meiotic sister chromatid segregation / Mis6-Sim4 complex / centromere complex assembly / establishment of meiotic sister chromatid cohesion ...Cbf1-Met4-Met28 complex / regulation of sulfur metabolic process / negative regulation of kinetochore assembly / negative regulation of meiotic DNA double-strand break formation involved in reciprocal meiotic recombination / COMA complex / maintenance of meiotic sister chromatid cohesion / meiotic sister chromatid segregation / Mis6-Sim4 complex / centromere complex assembly / establishment of meiotic sister chromatid cohesion / ascospore formation / attachment of spindle microtubules to kinetochore / centromeric DNA binding / CENP-A containing chromatin assembly / outer kinetochore / protein localization to chromosome, centromeric region / establishment of mitotic sister chromatid cohesion / kinetochore assembly / spindle pole body / protein localization to kinetochore / mitotic spindle assembly checkpoint signaling / chromosome, centromeric region / DNA replication initiation / meiotic cell cycle / chromosome segregation / kinetochore / DNA-binding transcription repressor activity, RNA polymerase II-specific / DNA-binding transcription activator activity, RNA polymerase II-specific / RNA polymerase II-specific DNA-binding transcription factor binding / protein dimerization activity / chromatin remodeling / RNA polymerase II cis-regulatory region sequence-specific DNA binding / DNA-binding transcription factor activity / cell division / protein-containing complex binding / regulation of transcription by RNA polymerase II / structural molecule activity / negative regulation of transcription by RNA polymerase II / positive regulation of transcription by RNA polymerase II / mitochondrion / nucleus / cytoplasm
Similarity search - Function
: / Kinetochore subunit Nkp2/Cnl2 / Cnl2/NKP2 family protein / : / : / CENPH protein / CENPU protein / Centromere protein O / Cenp-O kinetochore centromere component / Centromere protein Chl4/mis15/CENP-N ...: / Kinetochore subunit Nkp2/Cnl2 / Cnl2/NKP2 family protein / : / : / CENPH protein / CENPU protein / Centromere protein O / Cenp-O kinetochore centromere component / Centromere protein Chl4/mis15/CENP-N / Kinetochore protein CHL4 like / Helix-loop-helix DNA-binding domain / Helix-loop-helix DNA-binding domain superfamily / helix loop helix domain / Myc-type, basic helix-loop-helix (bHLH) domain / Myc-type, basic helix-loop-helix (bHLH) domain profile.
Similarity search - Domain/homology
Centromere-binding protein 1 / Inner kinetochore subunit IML3 / Inner kinetochore subunit AME1 / Inner kinetochore subunit CHL4 / Inner kinetochore subunit CNN1 / Inner kinetochore subunit MCM22 / Inner kinetochore subunit OKP1 / Inner kinetochore subunit CTF19 / Inner kinetochore subunit NKP2 / Inner kinetochore subunit MCM21 ...Centromere-binding protein 1 / Inner kinetochore subunit IML3 / Inner kinetochore subunit AME1 / Inner kinetochore subunit CHL4 / Inner kinetochore subunit CNN1 / Inner kinetochore subunit MCM22 / Inner kinetochore subunit OKP1 / Inner kinetochore subunit CTF19 / Inner kinetochore subunit NKP2 / Inner kinetochore subunit MCM21 / Inner kinetochore subunit MCM16 / Inner kinetochore subunit NKP1 / Inner kinetochore subunit CTF3 / Inner kinetochore subunit WIP1
Similarity search - Component
Biological speciesSaccharomyces cerevisiae (brewer's yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.4 Å
AuthorsDendooven TD / Zhang Z / Yang J / McLaughlin S / Schwabb J / Scheres S / Yatskevich S / Barford D
Funding support United Kingdom, Germany, 4 items
OrganizationGrant numberCountry
UK Research and Innovation (UKRI)MC_UP_1201/6 United Kingdom
Cancer Research UKC576/A14109 United Kingdom
UK Research and Innovation (UKRI)MC_UP_ A025_1013 United Kingdom
Boehringer Ingelheim Fonds (BIF) Germany
CitationJournal: Sci Adv / Year: 2023
Title: Cryo-EM structure of the complete inner kinetochore of the budding yeast point centromere.
Authors: Tom Dendooven / Ziguo Zhang / Jing Yang / Stephen H McLaughlin / Johannes Schwab / Sjors H W Scheres / Stanislau Yatskevich / David Barford /
Abstract: The point centromere of budding yeast specifies assembly of the large kinetochore complex to mediate chromatid segregation. Kinetochores comprise the centromere-associated inner kinetochore (CCAN) ...The point centromere of budding yeast specifies assembly of the large kinetochore complex to mediate chromatid segregation. Kinetochores comprise the centromere-associated inner kinetochore (CCAN) complex and the microtubule-binding outer kinetochore KNL1-MIS12-NDC80 (KMN) network. The budding yeast inner kinetochore also contains the DNA binding centromere-binding factor 1 (CBF1) and CBF3 complexes. We determined the cryo-electron microscopy structure of the yeast inner kinetochore assembled onto the centromere-specific centromere protein A nucleosomes (CENP-A). This revealed a central CENP-A with extensively unwrapped DNA ends. These free DNA duplexes bind two CCAN protomers, one of which entraps DNA topologically, positioned on the centromere DNA element I (CDEI) motif by CBF1. The two CCAN protomers are linked through CBF3 forming an arch-like configuration. With a structural mechanism for how CENP-A can also be linked to KMN involving only CENP-QU, we present a model for inner kinetochore assembly onto a point centromere and how it organizes the outer kinetochore for chromosome attachment to the mitotic spindle.
History
DepositionApr 26, 2023-
Header (metadata) releaseAug 9, 2023-
Map releaseAug 9, 2023-
UpdateAug 9, 2023-
Current statusAug 9, 2023Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_17224.png

Downloads & links

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Map

FileDownload / File: emd_17224.map.gz / Format: CCP4 / Size: 83.7 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
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Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.17 Å/pix.
x 280 pix.
= 327.88 Å		1.17 Å/pix.
x 280 pix.
= 327.88 Å		1.17 Å/pix.
x 280 pix.
= 327.88 Å

Surface

Projections

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Images are generated by Spider.

Voxel sizeX=Y=Z: 1.171 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.0354
Minimum - Maximum-0.089639254 - 0.14831993
Average (Standard dev.)0.00042256998 (±0.005386486)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions280280280
Spacing280280280
CellA=B=C: 327.88 Å
α=β=γ: 90.0 °

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Supplemental data

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Additional map: #1

Fileemd_17224_additional_1.map
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Half map: #2

Fileemd_17224_half_map_1.map
Projections & Slices
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Half map: #1

Fileemd_17224_half_map_2.map
Projections & Slices
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Sample components

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Entire : A complex of CBF1-CCAN bound to centromeric C0N3 DNA

EntireName: A complex of CBF1-CCAN bound to centromeric C0N3 DNA
Components
  • Complex: A complex of CBF1-CCAN bound to centromeric C0N3 DNA
    • Protein or peptide: Centromere-binding protein 1
    • DNA: C0N3 DNA
    • Protein or peptide: Inner kinetochore subunit MCM16
    • Protein or peptide: Inner kinetochore subunit CTF3
    • Protein or peptide: Inner kinetochore subunit MCM22
    • Protein or peptide: Inner kinetochore subunit IML3
    • Protein or peptide: Inner kinetochore subunit CHL4
    • Protein or peptide: Inner kinetochore subunit MCM21
    • Protein or peptide: Inner kinetochore subunit CTF19
    • Protein or peptide: Inner kinetochore subunit OKP1
    • Protein or peptide: Inner kinetochore subunit CNN1
    • Protein or peptide: Inner kinetochore subunit AME1
    • Protein or peptide: Inner kinetochore subunit WIP1
    • Protein or peptide: Inner kinetochore subunit NKP1
    • Protein or peptide: Inner kinetochore subunit NKP2
    • DNA: C0N3 DNA

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Supramolecule #1: A complex of CBF1-CCAN bound to centromeric C0N3 DNA

SupramoleculeName: A complex of CBF1-CCAN bound to centromeric C0N3 DNA / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#16
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)

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Macromolecule #1: Centromere-binding protein 1

MacromoleculeName: Centromere-binding protein 1 / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 39.444715 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MNSLANNNKL STEDEEIHSA RKRGYNEEQN YSEARKKQRD QGLLSQESND GNIDSALLSE GATLKGTQSQ YESGLTSNKD EKGSDDEDA SVAEAAVAAT VNYTDLIQGQ EDSSDAHTSN QTNANGEHKD SLNGERAITP SNEGVKPNTS LEGMTSSPME S TQQSKNDM ...String:
MNSLANNNKL STEDEEIHSA RKRGYNEEQN YSEARKKQRD QGLLSQESND GNIDSALLSE GATLKGTQSQ YESGLTSNKD EKGSDDEDA SVAEAAVAAT VNYTDLIQGQ EDSSDAHTSN QTNANGEHKD SLNGERAITP SNEGVKPNTS LEGMTSSPME S TQQSKNDM LIPLAEHDRG PEHQQDDEDN DDADIDLKKD ISMQPGRRGR KPTTLATTDE WKKQRKDSHK EVERRRRENI NT AINVLSD LLPVRESSKA AILACAAEYI QKLKETDEAN IEKWTLQKLL SEQNASQLAS ANEKLQEELG NAYKEIEYMK RVL RKEGIE YEDMHTHKKQ ENERKSTRSD NPHEA

UniProtKB: Centromere-binding protein 1

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Macromolecule #3: Inner kinetochore subunit MCM16

MacromoleculeName: Inner kinetochore subunit MCM16 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 21.1661 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString:
MTNSSEKQWE RIQQLEKEHV EVYRELLITL DRLYLIRKHN HAVILSHTQQ RLLEIRHQLQ INLEKTALLI RLLEKPDNTN VLFTKLQNL LEESNSLDYE LLQSLGAQSS LHKQLIESRA ERDELMSKLI ELSSKFPKPT IPPDDSDTAG KQVEVEKENE T IQELMIAL QIHSGYTNIS YTI

UniProtKB: Inner kinetochore subunit MCM16

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Macromolecule #4: Inner kinetochore subunit CTF3

MacromoleculeName: Inner kinetochore subunit CTF3 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 84.345633 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MSLILDDIIL SLTNANERTP PQALKTTLSL LYEKSKQYGL SSPQLQALVR LLCETSIIDT VTKVYIVENC FLPDGYLTKE LLLEIINHL GTPTVFSRYR IQTPPVLQSA LCKWLVHVYF LFPVHSEREH NISSSIWLHL WQFSFLQKWI TPLVIWQATT P VDVKPWKL ...String:
MSLILDDIIL SLTNANERTP PQALKTTLSL LYEKSKQYGL SSPQLQALVR LLCETSIIDT VTKVYIVENC FLPDGYLTKE LLLEIINHL GTPTVFSRYR IQTPPVLQSA LCKWLVHVYF LFPVHSEREH NISSSIWLHL WQFSFLQKWI TPLVIWQATT P VDVKPWKL SIIKRCAMHP GYRDAPGSAT LILQRFQCLV GASSQITESI ITINCNRKTL KSHRNLKLDA HFLSILKRIL SR AHPANFP ADTVQNTIDM YLSEIHQLGA DSIYPLRLQS LPEYVPSDST VSLWDVTSLE QLAQNWPQLH IPNDVDYMMK PSL NSNVLL PRKVMSRDSL KHLYSSIILI KNSRDESSSP YEWCIWQLKR CFAHQIETPQ EVIPIIISVS SMDNKLSSRI IQTF CNLKY LKLDELTLKK VCGGILPLWK PELISGTREF FVKFMASIFM WSTRDGHDNN CTFSETCFYV LQMITNWVLD DKLIA LGLT LLHDMQSLLT LDKIFNNATS NRFSTMAFIS SLDILTQLSK QTKSDYAIQY LIVGPDIMNK VFSSDDPLLL SAACRY LVA TKNKLMQYPS TNKFVRMQNQ YIMDLTNYLY RNKVLSSKSL FGVSPDFFKQ ILENLYIPTA DFKNAKFFTI TGIPALS YI CIIILRRLET AENTKIKFTS GIINEETFNN FFRVHHDEIG QHGWIKGVNN IHDLRVKILM HLSNTANPYR DIAAFLFT Y LKSLSKYSVQ NS

UniProtKB: Inner kinetochore subunit CTF3

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Macromolecule #5: Inner kinetochore subunit MCM22

MacromoleculeName: Inner kinetochore subunit MCM22 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 27.602541 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MDVEKDVLDV YIKNLENQIG NKRYFLKQAQ GAIDEITKRS LDTEGKPVNS EVFTELLRKP MFFSERADPI GFSLTSNFLS LRAQSSSEW LSLMNDQSVD QKAMLLLQNN INSDLKELLR KLQHQMTIMD SKKQDHAHIR TRKARNKELW DSLADFLKGY L VPNLDDND ...String:
MDVEKDVLDV YIKNLENQIG NKRYFLKQAQ GAIDEITKRS LDTEGKPVNS EVFTELLRKP MFFSERADPI GFSLTSNFLS LRAQSSSEW LSLMNDQSVD QKAMLLLQNN INSDLKELLR KLQHQMTIMD SKKQDHAHIR TRKARNKELW DSLADFLKGY L VPNLDDND ESIDSLTNEV MLLMKRLIEH DLNLTLNDFS SKTIPIYRLL LRANIITVIE GSTNPGTKYI KLIDFNETSL T

UniProtKB: Inner kinetochore subunit MCM22

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Macromolecule #6: Inner kinetochore subunit IML3

MacromoleculeName: Inner kinetochore subunit IML3 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 28.093223 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MPYTWKFLGI SKQLSLENGI AKLNQLLNLE VDLDIQTIRV PSDPDGGTAA DEYIRYEMRL DISNLDEGTY SKFIFLGNSK MEVPMFLCY CGTDNRNEVV LQWLKAEYGV IMWPIKFEQK TMIKLADASI VHVTKENIEQ ITWFSSKLYF EPETQDKNLR Q FSIEIPRE ...String:
MPYTWKFLGI SKQLSLENGI AKLNQLLNLE VDLDIQTIRV PSDPDGGTAA DEYIRYEMRL DISNLDEGTY SKFIFLGNSK MEVPMFLCY CGTDNRNEVV LQWLKAEYGV IMWPIKFEQK TMIKLADASI VHVTKENIEQ ITWFSSKLYF EPETQDKNLR Q FSIEIPRE SCEGLALGYG NTMHPYNDAI VPYIYNETGM AVERLPLTSV ILAGHTKIMR ESIVTSTRSL RNRVLAVVLQ SI QFTSE

UniProtKB: Inner kinetochore subunit IML3

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Macromolecule #7: Inner kinetochore subunit CHL4

MacromoleculeName: Inner kinetochore subunit CHL4 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 52.743723 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MSNELRLEDN YVPTSDTLVV FKQLMKLPVT VLYDLTLSWF AKFGGSFDGD IYLLTETLDL LIEKGVRRNV IVNRILYVYW PDGLNVFQL AEIDCHLMIS KPEKFKWLPS KALRGDGKPY VVKLQPAKFI ENLQTDLAKI YHCHVYMFKH PSLPVLITRI Q LFDSNNLF ...String:
MSNELRLEDN YVPTSDTLVV FKQLMKLPVT VLYDLTLSWF AKFGGSFDGD IYLLTETLDL LIEKGVRRNV IVNRILYVYW PDGLNVFQL AEIDCHLMIS KPEKFKWLPS KALRGDGKPY VVKLQPAKFI ENLQTDLAKI YHCHVYMFKH PSLPVLITRI Q LFDSNNLF LSTPNIGSIN KESLYNKLDK FQGKPLISRR PYYVAFPLNS PIIFHSVDKD IYARLVLQSI SRTISERETI IF KPVQKIP VKSIHNIMTL LGPSRFAESM GPWECYASAN FERSPLHDYK KHQGLTGKKV MVREFDDSFL NDDENFYGKE EPE IRRLRL EKNMIKFKGS ANGVMDQKYN DLKEFNEHVH NIRNGKKNED SGEPVYISRY SSLVPIEKVG FTLKNEINSR IITI KLKFN GNDIFGGLHE LCDKNLINID KVPGWLAGEN GSFSGTIMNG DFQREQVAKG GLL

UniProtKB: Inner kinetochore subunit CHL4

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Macromolecule #8: Inner kinetochore subunit MCM21

MacromoleculeName: Inner kinetochore subunit MCM21 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 43.028879 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MSRIDDLQQD IESLLSEINS LEESREKLKA KIKDKRKNEE SANPIVQEFE DLFDQFPQLN NFLFNEHPEL EETDDKDISR AQADIPATP IPYEPKKRAK LENEEILPEQ EWVLKTQPMV QHQMFDPGVA DLLDTDILTS PSKRKRKLKI DDISTSDRSE L EDYIVLEN ...String:
MSRIDDLQQD IESLLSEINS LEESREKLKA KIKDKRKNEE SANPIVQEFE DLFDQFPQLN NFLFNEHPEL EETDDKDISR AQADIPATP IPYEPKKRAK LENEEILPEQ EWVLKTQPMV QHQMFDPGVA DLLDTDILTS PSKRKRKLKI DDISTSDRSE L EDYIVLEN VYRMFGITFF PLVDPIDLKI KDASGEIFVD REMLGIRLEV FSERTSQFEK PHYVLLKKRI KSNSWFLFKH TI PSFIDVQ GIFDDTNGGL VISHDDAYLF AKRVFLQLVE VQKRRQIFKD LEAKKIIHDL DLDLESSMVS FFVKDIKVEL FVK QNEIVS CSILDDIHDF SQNNKSKWEI ALLGSLDDLE LKLNHSFATI FK

UniProtKB: Inner kinetochore subunit MCM21

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Macromolecule #9: Inner kinetochore subunit CTF19

MacromoleculeName: Inner kinetochore subunit CTF19 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 42.841113 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MDFTSDTTNS HDTSNSHLSL EDAVGTHHAG EADVNIDGDE KQQLSLLDDD QVRALKLQEE KDALLTRRNT LLQEIQTYQN ILMKENNSK TKNGDILQND ITQDFLNLIS ISSSNPNSAI SDRKRVERIN GLTNLQKELV TKYDTLPLLN MNLRLSYLRD H TYPHLQVS ...String:
MDFTSDTTNS HDTSNSHLSL EDAVGTHHAG EADVNIDGDE KQQLSLLDDD QVRALKLQEE KDALLTRRNT LLQEIQTYQN ILMKENNSK TKNGDILQND ITQDFLNLIS ISSSNPNSAI SDRKRVERIN GLTNLQKELV TKYDTLPLLN MNLRLSYLRD H TYPHLQVS VQSRDRVHND GIEVLVVNYK FCRNTMNPFE IQFKMFYKFE DSTLLKWEIL RISTNVRLKA KQLLATRNFQ KC LLSLYEF DKIKSKKTGI FQNLINLLKR KTRCYLMNNS DSLIVERVIR EGRLTTIKLQ INFIITMPGE RGKPRNCFLP MSK ISIALW KGGERFNQID LDEICYGLIK EYGVKTGLKE ICNVCLFPDM YAR

UniProtKB: Inner kinetochore subunit CTF19

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Macromolecule #10: Inner kinetochore subunit OKP1

MacromoleculeName: Inner kinetochore subunit OKP1 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 47.427246 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MAADRDNFLQ NIENDSINNG QAMDLSPNRS SSESDSSILM NVNDIKTLRL DVAPEAKSTQ SKKSLFYENS DDAEEGEIEE RTNKEEGQY HHKGSKQLRF EVGKESTGKL QSHLSDGSAT SGEGNVRPWE FRKVIQAEYR ERLPRNYELK HWKKPSKIMI G SILRLLET ...String:
MAADRDNFLQ NIENDSINNG QAMDLSPNRS SSESDSSILM NVNDIKTLRL DVAPEAKSTQ SKKSLFYENS DDAEEGEIEE RTNKEEGQY HHKGSKQLRF EVGKESTGKL QSHLSDGSAT SGEGNVRPWE FRKVIQAEYR ERLPRNYELK HWKKPSKIMI G SILRLLET NTVSALDSVF EKYEKEMNQM THGDNNEVKR IYSKKERLLE IILTKIKKKL RQAKFPSRIS ERDLDIEYIY SK RQFIQNR YSQELQNNER LEAILSREQN LLEETRKLCM NLKTNNKKRL TEKLIQKDLH PVLNKAMEYT YGLESTNGFM HPD GPVTFR NDSHELNLML NDPIKSTADV RLDKEEVLSL LPSLKEYTKK SKELKETMGQ MISDSHEEEI KEVFVPHHES HQDK TEEDI H

UniProtKB: Inner kinetochore subunit OKP1

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Macromolecule #11: Inner kinetochore subunit CNN1

MacromoleculeName: Inner kinetochore subunit CNN1 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 41.359785 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MSTPRKAAGN NENTEVSEIR TPFRERALEE QRLKDEVLIR NTPGYRKLLS ASTKSHDILN KDPNEVRSFL QDLSQVLARK SQGNDTTTN KTQARNLIDE LAYEESQPEE NELLRSRSEK LTDNNIGNET QPDYTSLSQT VFAKLQERDK GLKSRKIDPI I IQDVPTTG ...String:
MSTPRKAAGN NENTEVSEIR TPFRERALEE QRLKDEVLIR NTPGYRKLLS ASTKSHDILN KDPNEVRSFL QDLSQVLARK SQGNDTTTN KTQARNLIDE LAYEESQPEE NELLRSRSEK LTDNNIGNET QPDYTSLSQT VFAKLQERDK GLKSRKIDPI I IQDVPTTG HEDELTVHSP DKANSISMEV LRTSPSIGMD QVDEPPVRDP VPISITQQEE PLSEDLPSDD KEETEEAENE DY SFENTSD ENLDDIGNDP IRLNVPAVRR SSIKPLQIMD LKHLTRQFLN ENRIILPKQT WSTIQEESLN IMDFLKQKIG TLQ KQELVD SFIDMGIINN VDDMFELAHE LLPLELQSRI ESYLF

UniProtKB: Inner kinetochore subunit CNN1

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Macromolecule #12: Inner kinetochore subunit AME1

MacromoleculeName: Inner kinetochore subunit AME1 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 37.506723 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MDRDTKLAFR LRGSHSRRTD DIDDDVIVFK TPNAVYREEN SPIQSPVQPI LSSPKLANSF EFPITTNNVN AQDRHEHGYQ PLDAEDYPM IDSENKSLIS ESPQNVRNDE DLTTRYNFDD IPIRQLSSSI TSVTTIDVLS SLFINLFEND LIPQALKDFN K SDDDQFRK ...String:
MDRDTKLAFR LRGSHSRRTD DIDDDVIVFK TPNAVYREEN SPIQSPVQPI LSSPKLANSF EFPITTNNVN AQDRHEHGYQ PLDAEDYPM IDSENKSLIS ESPQNVRNDE DLTTRYNFDD IPIRQLSSSI TSVTTIDVLS SLFINLFEND LIPQALKDFN K SDDDQFRK LLYKLDLRLF QTISDQMTRD LKDILDINVS NNELCYQLKQ VLARKEDLNQ QIISVRNEIQ ELKAGKDWHD LQ NEQAKLN DKVKLNKRLN DLTSTLLGKY EGDRKIMSQD SEDDSIRDDS NILDIAHFVD LMDPYNGLLK KINKINENLS NEL QPSL

UniProtKB: Inner kinetochore subunit AME1

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Macromolecule #13: Inner kinetochore subunit WIP1

MacromoleculeName: Inner kinetochore subunit WIP1 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 10.255458 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString:
MDTEALANYL LRQLSLDAEE NKLEDLLQRQ NEDQESSQEY NKKLLLACGF QAILRKILLD ARTRATAEGL REVYPYHIEA ATQAFLDSQ

UniProtKB: Inner kinetochore subunit WIP1

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Macromolecule #14: Inner kinetochore subunit NKP1

MacromoleculeName: Inner kinetochore subunit NKP1 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 27.006451 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString: MTDTYNSISN FIENELTALL SSDDYLMDDL AGELPNEVCR LLKAQVIEKR KDAMSRGKQD LLSKEIYDNE SELRASQSQQ IMELVGDIP KYSLGSELRN RVEGEPQSTS IERLIEDVLK LPQMEVADEE EVEVENDLKV LSEYSNLRKD LILKCQALQI G ESKLSDIL ...String:
MTDTYNSISN FIENELTALL SSDDYLMDDL AGELPNEVCR LLKAQVIEKR KDAMSRGKQD LLSKEIYDNE SELRASQSQQ IMELVGDIP KYSLGSELRN RVEGEPQSTS IERLIEDVLK LPQMEVADEE EVEVENDLKV LSEYSNLRKD LILKCQALQI G ESKLSDIL SQTNSINSLT TSIKEASEDD DISEYFATYN GKLVVALEEM KLLLEEAVKT FGNSPEKREK IKKILSELKK

UniProtKB: Inner kinetochore subunit NKP1

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Macromolecule #15: Inner kinetochore subunit NKP2

MacromoleculeName: Inner kinetochore subunit NKP2 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 17.877033 KDa
Recombinant expressionOrganism: Trichoplusia ni (cabbage looper)
SequenceString:
MNSEQLLHNY VSDSLLTTLI SFQEFKQQLQ SYTSDEQQLQ HWYELLQARD ARVTSELEAR IKQFFITLRS RLLRFLESEQ LSHSLSLET LIDALYKIND LLQQRLQILD DAIQEKTSEL AEFENMVRSP SAGDNAIPGL LQIIQSYINL LEEN

UniProtKB: Inner kinetochore subunit NKP2

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Macromolecule #2: C0N3 DNA

MacromoleculeName: C0N3 DNA / type: dna / ID: 2 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 47.194199 KDa
SequenceString: (DT)(DT)(DC)(DA)(DA)(DT)(DG)(DA)(DA)(DA) (DT)(DA)(DT)(DA)(DT)(DA)(DT)(DT)(DT)(DC) (DT)(DT)(DA)(DC)(DT)(DA)(DT)(DT)(DT) (DC)(DT)(DT)(DT)(DT)(DT)(DT)(DA)(DA)(DC) (DT) (DT)(DT)(DC)(DG)(DG)(DA) ...String:
(DT)(DT)(DC)(DA)(DA)(DT)(DG)(DA)(DA)(DA) (DT)(DA)(DT)(DA)(DT)(DA)(DT)(DT)(DT)(DC) (DT)(DT)(DA)(DC)(DT)(DA)(DT)(DT)(DT) (DC)(DT)(DT)(DT)(DT)(DT)(DT)(DA)(DA)(DC) (DT) (DT)(DT)(DC)(DG)(DG)(DA)(DA)(DA) (DT)(DC)(DA)(DA)(DA)(DT)(DA)(DC)(DA)(DC) (DT)(DA) (DA)(DT)(DA)(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)(DA) (DT)(DG)(DT)(DG)(DA)(DC)(DT)(DT)(DA)(DT)

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Macromolecule #16: C0N3 DNA

MacromoleculeName: C0N3 DNA / type: dna / ID: 16 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 47.239277 KDa
SequenceString: (DA)(DT)(DA)(DA)(DG)(DT)(DC)(DA)(DC)(DA) (DT)(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:
(DA)(DT)(DA)(DA)(DG)(DT)(DC)(DA)(DC)(DA) (DT)(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)(DT)(DA)(DT)(DT)(DA)(DG) (DT)(DG)(DT)(DA) (DT)(DT)(DT)(DG)(DA) (DT)(DT)(DT)(DC)(DC)(DG)(DA)(DA)(DA)(DG) (DT)(DT)(DA)(DA)(DA) (DA)(DA)(DA)(DG) (DA)(DA)(DA)(DT)(DA)(DG)(DT)(DA)(DA)(DG) (DA)(DA)(DA)(DT)(DA)(DT) (DA)(DT)(DA) (DT)(DT)(DT)(DC)(DA)(DT)(DT)(DG)(DA)(DA)

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

BufferpH: 8
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 2.6 µm / Nominal defocus min: 1.0 µm
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 40.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: NONE
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.4 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 43467

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