title: CENP-C is a blueprint for constitutive centromere-associated network assembly within human kinetochores.
abstract: Kinetochores are multisubunit complexes that assemble on centromeres to bind spindle microtubules and promote faithful chromosome segregation during cell division. A 16-subunit complex named the constitutive centromere-associated network (CCAN) creates the centromere-kinetochore interface. CENP-C, a CCAN subunit, is crucial for kinetochore assembly because it links centromeres with the microtubule-binding interface of kinetochores. The role of CENP-C in CCAN organization, on the other hand, had been incompletely understood. In this paper, we combined biochemical reconstitution and cellular investigations to unveil how CENP-C promotes kinetochore targeting of other CCAN subunits. The so-called PEST domain in the N-terminal half of CENP-C interacted directly with the four-subunit CCAN subcomplex CENP-HIKM. We identified crucial determinants of this interaction whose mutation prevented kinetochore localization of CENP-HIKM and of CENP-TW, another CCAN subcomplex. When considered together with previous observations, our data point to CENP-C as a blueprint for kinetochore assembly.
authors.affiliation: Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, 44227 Dortmund, Germany.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Klare
authors.affiliation: Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, 44227 Dortmund, Germany.
authors.forename: John R
authors.initials: JR
authors.lastname: Weir
authors.affiliation: Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, 44227 Dortmund, Germany Department of Experimental Oncology, European Institute of Oncology, 20139 Milan, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Basilico
authors.affiliation: Gene Center Munich, Ludwig-Maximilians-Universität München, 81377 Munich, Germany.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Zimniak
authors.affiliation: Department of Experimental Oncology, European Institute of Oncology, 20139 Milan, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Massimiliano
authors.affiliation: Department of Structural Biochemistry, Max Planck Institute of Molecular Physiology, 44227 Dortmund, Germany.
authors.forename: Nina
authors.initials: N
authors.lastname: Ludwigs
authors.affiliation: Gene Center Munich, Ludwig-Maximilians-Universität München, 81377 Munich, Germany.
authors.forename: Franz
authors.initials: F
authors.lastname: Herzog
authors.affiliation: Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, 44227 Dortmund, Germany Centre for Medical Biotechnology, Faculty of Biology, University Duisburg-Essen, 45141 Essen, Germany andrea.musacchio@mpi-dortmund.mpg.de.
authors.forename: Andrea
authors.initials: A
authors.lastname: Musacchio
mesh_terms: D002868:Chromosomal Proteins, Non-Histone / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D008969:Molecular Sequence Data; D060066:Protein Interaction Maps; D021381:Protein Transport
chemical_list: D002868:Chromosomal Proteins, Non-Histone; C060905:centromere protein C