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TitleCryo-EM structure of the complete inner kinetochore of the budding yeast point centromere.
Journal, issue, pagesSci Adv, Vol. 9, Issue 30, Page eadg7480, Year 2023
Publish dateJul 28, 2023
AuthorsTom Dendooven / Ziguo Zhang / Jing Yang / Stephen H McLaughlin / Johannes Schwab / Sjors H W Scheres / Stanislau Yatskevich / David Barford /
PubMed AbstractThe 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.
External linksSci Adv / PubMed:37506202 / PubMed Central
MethodsEM (single particle)
Resolution3.4 - 4.9 Å
Structure data

17224

8ovw

EMDB-17224, PDB-8ovw:
Cryo-EM structure of CBF1-CCAN bound topologically to centromeric DNA
Method: EM (single particle) / Resolution: 3.4 Å

17225

8ovx

EMDB-17225, PDB-8ovx:
Cryo-EM structure of yeast CENP-OPQU+ bound to the CENP-A N-terminus
Method: EM (single particle) / Resolution: 3.4 Å

17226

8ow0

EMDB-17226, PDB-8ow0:
Cryo-EM structure of CBF1-CCAN bound topologically to a centromeric CENP-A nucleosome
Method: EM (single particle) / Resolution: 3.4 Å

17227

8ow1

EMDB-17227, PDB-8ow1:
Cryo-EM structure of the yeast Inner kinetochore bound to a CENP-A nucleosome.
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-17368: Multibody map of the CENP-A nucleosome as part of the inner kinetochore
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-17371: Multibody map of CCAN(Topo) bound to CON3 DNA as part of the Inner kinetochore.
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-17372: Multibody map of CCAN(Non-Topo) bound to C0N3 DNA as part of the inner kinetochore.
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-17374: Multibody map of CBF3:CENP-HIK as part of the inner kinetochore
Method: EM (single particle) / Resolution: 3.7 Å

EMDB-17376: Consensus map of the yeast inner kinetochore
Method: EM (single particle) / Resolution: 4.9 Å

Source
  • saccharomyces cerevisiae (brewer's yeast)
KeywordsCELL CYCLE / kinetochore / point centromere / CENP-A nucleosome / topological entrapment / centromeric DNA

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