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Cited by 58Microtubule and mitosis dynamicsGenomics and Chromatin DynamicsNuclear Structure and Function

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Cryo-EM structure of the complete inner kinetochore of the budding yeast point centromere

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Dendooven, Tom, Zhang, Ziguo, Yang, Jing, McLaughlin, Stephen H., Schwab, Johannes, Scheres, Sjors H. W., Yatskevich, Stanislau, Barford, David·2023·Science Advances
DOI 10.1126/sciadv.adg7480PMC PMC10381965

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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) 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 Nuc ). This revealed a central CENP-A Nuc 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 Nuc 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.

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Metadata

Authors
Dendooven, T., Zhang, Z., Yang, J., McLaughlin, S. H., Schwab, J., Scheres, S. H. W., Yatskevich, S., & Barford, D.
Affiliations
MRC Laboratory of Molecular Biology (GB)
Corresponding author
Barford, David
ORCID
Venue
Science Advances
Publisher
American Association for the Advancement of Science (AAAS)
Published
2023-07-28
Volume
9
Issue
30
Pages
eadg7480-eadg7480
ISSN
2375-2548 (ISSN-L)
DOI
10.1126/sciadv.adg7480
PMC
PMC10381965
OpenAlex
W4385334888
Cited by
58
References
5
Citations by year
2026: 16 · 2025: 26 · 2024: 11 · 2023: 5
FWCI
8.36
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Science Advances (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • Wellcome Trust
  • Diamond Light Source (BI31336)
  • Alan Turing Institute (EP/W006022/1)
  • UK Research and Innovation (MC_UP_1201/6, EP/W006022/1)
  • Cancer Research UK (24307, C576/A14109, 14109)
  • Medical Research Council (MC_UP_1201/6)
  • Engineering and Physical Sciences Research Council (EP/W006022/1)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
Keywords
inner kinetochore, Saccharomyces cerevisiae, CBF1, CBF3, KMN network, kinetochore assembly, chromosome segregation
MeSH terms
Centromere Protein A, Centromere, Chromosomal Proteins, Non-Histone, DNA, Saccharomycetales, Nuclear Proteins, Saccharomyces cerevisiae, Kinetochores, Cryoelectron Microscopy, Protein Subunits, Saccharomyces cerevisiae Proteins, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors

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