CENP-T provides a structural platform for outer kinetochore assembly

EMBO J. 2013 Feb 6;32(3):424-36. doi: 10.1038/emboj.2012.348. Epub 2013 Jan 18.

Abstract

The kinetochore forms a dynamic interface with microtubules from the mitotic spindle during mitosis. The Ndc80 complex acts as the key microtubule-binding complex at kinetochores. However, it is unclear how the Ndc80 complex associates with the inner kinetochore proteins that assemble upon centromeric chromatin. Here, based on a high-resolution structural analysis, we demonstrate that the N-terminal region of vertebrate CENP-T interacts with the 'RWD' domain in the Spc24/25 portion of the Ndc80 complex. Phosphorylation of CENP-T strengthens a cryptic hydrophobic interaction between CENP-T and Spc25 resulting in a phospho-regulated interaction that occurs without direct recognition of the phosphorylated residue. The Ndc80 complex interacts with both CENP-T and the Mis12 complex, but we find that these interactions are mutually exclusive, supporting a model in which two distinct pathways target the Ndc80 complex to kinetochores. Our results provide a model for how the multiple protein complexes at kinetochores associate in a phospho-regulated manner.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Calorimetry
  • Cell Line, Tumor
  • Chickens
  • Chromatography, Gel
  • Chromosomal Proteins, Non-Histone / chemistry
  • Chromosomal Proteins, Non-Histone / metabolism*
  • Crystallization
  • Cytoskeletal Proteins
  • Humans
  • Kinetochores / chemistry
  • Kinetochores / metabolism*
  • Microscopy, Fluorescence
  • Microtubule-Associated Proteins / chemistry
  • Microtubule-Associated Proteins / metabolism*
  • Models, Molecular*
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / metabolism*
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / metabolism
  • Phosphorylation
  • Species Specificity

Substances

  • CENPT protein, human
  • Chromosomal Proteins, Non-Histone
  • Cytoskeletal Proteins
  • MIS12 protein, human
  • Microtubule-Associated Proteins
  • Multiprotein Complexes
  • NDC80 protein, human
  • Nuclear Proteins
  • SPC24 protein, human
  • SPC25 protein, human

Associated data

  • PDB/3VZ9
  • PDB/3VZA