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Updated: May 27, 2026
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Protein Purification Technique that Allows Detection of Sumoylation and Ubiquitination of Budding Yeast Kinetochore Proteins Ndc10 and Ndc80
Published on: May 3, 2015
Ndc10 is a platform for inner kinetochore assembly in budding yeast
Uhn-Soo Cho1, Stephen C Harrison
1Jack and Eileen Connors Structural Biology Laboratory and Howard Hughes Medical Institute, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, USA.
Nature Structural & Molecular Biology
|December 6, 2011
Summary
The Ndc10 protein in budding yeast forms a DNA loop at centromeres, acting as a platform for inner kinetochore assembly to ensure accurate chromosome segregation during cell division.
Area of Science:
- Molecular Biology
- Cell Biology
- Structural Biology
Background:
- Kinetochores are crucial for faithful chromosome segregation during mitosis.
- The CBF3 complex, including Ndc10, recognizes centromeric DNA in yeasts.
- Understanding Ndc10's role is key to kinetochore assembly.
Purpose of the Study:
- To elucidate the structural organization of Kluyveromyces lactis Ndc10.
- To determine the DNA-binding mechanism of Ndc10 domains I-II.
- To investigate Ndc10's interactions with other inner kinetochore proteins.
Main Methods:
- X-ray crystallography to determine the structure of Ndc10 domains I-II bound to DNA.
- Biochemical assays to analyze DNA binding and protein interactions.
- In vitro association experiments to map protein-protein interactions.
Main Results:
- The five-domain organization of Kluyveromyces lactis Ndc10 was described.
- The 2.8 Å structure of Ndc10 domains I-II bound to DNA revealed a resemblance to tyrosine DNA recombinases.
- Ndc10 dimer stabilizes a DNA loop by binding separate DNA fragments, and specific domains interact with inner kinetochore proteins.
Conclusions:
- Ndc10 plays a critical role in centromeric DNA organization.
- Ndc10 functions as a scaffold, facilitating the assembly of the inner kinetochore complex.
- This provides a structural basis for understanding kinetochore assembly and function in chromosome segregation.
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