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Nanoscale structural organization and stoichiometry of the budding yeast kinetochore

View ORCID ProfileKonstanty Cieslinski, View ORCID ProfileYu-Le Wu, Lisa Nechyporenko, Sarah Janice Hörner, Duccio Conti, View ORCID ProfileJonas Ries
doi: https://doi.org/10.1101/2021.12.01.469648
Konstanty Cieslinski
1European Molecular Biology Laboratory, Cell Biology and Biophysics Unit, Heidelberg, Germany
2DKFZ, Translational Radiation Oncology Unit, Heidelberg, Germany
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  • ORCID record for Konstanty Cieslinski
Yu-Le Wu
1European Molecular Biology Laboratory, Cell Biology and Biophysics Unit, Heidelberg, Germany
3Collaboration for joint PhD degree between EMBL and Heidelberg University, Faculty of Biosciences, Heidelberg, Germany
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Lisa Nechyporenko
1European Molecular Biology Laboratory, Cell Biology and Biophysics Unit, Heidelberg, Germany
4Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Heidelberg, Germany
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Sarah Janice Hörner
1European Molecular Biology Laboratory, Cell Biology and Biophysics Unit, Heidelberg, Germany
5Institute of Molecular and Cell Biology, Mannheim University of Applied Sciences and Interdisciplinary Center for Neuroscience, Heidelberg University, Heidelberg, Germany
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Duccio Conti
6Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
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Jonas Ries
1European Molecular Biology Laboratory, Cell Biology and Biophysics Unit, Heidelberg, Germany
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  • For correspondence: jonas.ries@embl.de
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Abstract

Proper chromosome segregation is crucial for cell division. In eukaryotes, this is achieved by the kinetochore, an evolutionarily conserved multi-protein complex that physically links the DNA to spindle microtubules, and takes an active role in monitoring and correcting erroneous spindle-chromosome attachments. Our mechanistic understanding of these functions, and how they ensure an error-free outcome of mitosis, is still limited, partly because we lack a comprehensive understanding of the kinetochore structure in the cell. In this study, we use single molecule localization microscopy to visualize individual kinetochore complexes in situ in budding yeast. For all major kinetochore proteins, we measured abundance and position within the metaphase kinetochore. Based on this comprehensive dataset, we propose a quantitative model of the budding yeast kinetochore. While confirming many aspects of previous reports based on bulk imaging of kinetochores, our results present a somewhat different but unifying model of the inner kinetochore. We find that the centromere-specialized histone Cse4 is present in more than two copies per kinetochore along with its binding partner Mif2.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • Spelling mistake in author list corrected.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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Posted December 03, 2021.
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Nanoscale structural organization and stoichiometry of the budding yeast kinetochore
Konstanty Cieslinski, Yu-Le Wu, Lisa Nechyporenko, Sarah Janice Hörner, Duccio Conti, Jonas Ries
bioRxiv 2021.12.01.469648; doi: https://doi.org/10.1101/2021.12.01.469648
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Nanoscale structural organization and stoichiometry of the budding yeast kinetochore
Konstanty Cieslinski, Yu-Le Wu, Lisa Nechyporenko, Sarah Janice Hörner, Duccio Conti, Jonas Ries
bioRxiv 2021.12.01.469648; doi: https://doi.org/10.1101/2021.12.01.469648

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