Ndc10 is a platform for inner kinetochore assembly in budding yeast.

Publication Type:

Journal Article

Authors:

Cho, Uhn-Soo; Harrison, Stephen C

Source:

Nat Struct Mol Biol, Volume 19, Issue 1, p.48-55 (2011)

Keywords:

Amino Acid Sequence, Binding Sites, Crystallography, X-Ray, DNA, Fungal, DNA-Binding Proteins, Electrophoresis, Polyacrylamide Gel, Electrophoretic Mobility Shift Assay, Fungal Proteins, Kinetochores, Kluyveromyces, Models, Molecular, Molecular Sequence Data, Mutation, Protein Binding, Protein Multimerization, Protein Structure, Tertiary, Sequence Homology, Amino Acid

Abstract:

<p>Kinetochores link centromeric DNA to spindle microtubules and ensure faithful chromosome segregation during mitosis. In point-centromere yeasts, the CBF3 complex Skp1-Ctf13-(Cep3)(2)-(Ndc10)(2) recognizes a conserved centromeric DNA element through contacts made by Cep3 and Ndc10. We describe here the five-domain organization of Kluyveromyces lactis Ndc10 and the structure at 2.8 Å resolution of domains I-II (residues 1-402) bound to DNA. The structure resembles tyrosine DNA recombinases, although it lacks both endonuclease and ligase activities. Structural and biochemical data demonstrate that each subunit of the Ndc10 dimer binds a separate fragment of DNA, suggesting that Ndc10 stabilizes a DNA loop at the centromere. We describe in vitro association experiments showing that specific domains of Ndc10 interact with each of the known inner-kinetochore proteins or protein complexes in budding yeast. We propose that Ndc10 provides a central platform for inner-kinetochore assembly.</p>