title: CENP-T provides a structural platform for outer kinetochore assembly.
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.
authors: Department of Molecular Genetics, National Institute of Genetics and The Graduate University for Advanced Studies SOKENDAI, Shizuoka 411-8540, Japan.
authors: Tatsuya
authors: T
authors: Nishino
authors: Florencia
authors: F
authors: Rago
authors: Tetsuya
authors: T
authors: Hori
authors: Kentaro
authors: K
authors: Tomii
authors: Iain M
authors: IM
authors: Cheeseman
authors: Tatsuo
authors: T
authors: Fukagawa
mesh_terms: D000818:Animals; D002151:Calorimetry; D045744:Cell Line, Tumor; D002645:Chickens; D002850:Chromatography, Gel; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000378:metabolism*; D003460:Crystallization; D003598:Cytoskeletal Proteins; D006801:Humans; D018386:Kinetochores / Q000737:chemistry / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry / Q000378:metabolism*; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D013045:Species Specificity
chemical_list: C534001:CENPT protein, human; D002868:Chromosomal Proteins, Non-Histone; D003598:Cytoskeletal Proteins; C492664:MIS12 protein, human; D008869:Microtubule-Associated Proteins; D046912:Multiprotein Complexes; C120529:NDC80 protein, human; D009687:Nuclear Proteins; C484695:SPC24 protein, human; C484694:SPC25 protein, human