Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany; Department of Experimental Oncology, European Institute of Oncology, Milan, Italy
Stefano Maffini
Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
John R Weir
Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
Daniel Prumbaum
Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Dortmund, Germany
Ana M Rojas
Computational Biology and Bioinformatics Group, Institute of Biomedicine of Seville, Campus Hospital Universitario Virgen del Rocio, Seville, Spain
Tomasz Zimniak
Department of Biochemistry and Gene Center, Ludwig-Maximilians-Universität, München, Munich, Germany
Anna De Antoni
Department of Experimental Oncology, European Institute of Oncology, Milan, Italy
Sadasivam Jeganathan
Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
Beate Voss
Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
Suzan van Gerwen
Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
Veronica Krenn
Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany; Department of Experimental Oncology, European Institute of Oncology, Milan, Italy
Lucia Massimiliano
Department of Experimental Oncology, European Institute of Oncology, Milan, Italy
Alfonso Valencia
Structural Biology and Biocomputing Programme, Spanish National Cancer Centre–CNIO, Madrid, Spain
Ingrid R Vetter
Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany
Franz Herzog
Department of Biochemistry and Gene Center, Ludwig-Maximilians-Universität, München, Munich, Germany
Stefan Raunser
Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Dortmund, Germany
Sebastiano Pasqualato
Department of Experimental Oncology, European Institute of Oncology, Milan, Italy
Andrea Musacchio
Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Dortmund, Germany; Centre for Medical Biotechnology, Faculty of Biology, University of Duisburg-Essen, Essen, Germany
Kinetochores, multi-subunit complexes that assemble at the interface with centromeres, bind spindle microtubules to ensure faithful delivery of chromosomes during cell division. The configuration and function of the kinetochore–centromere interface is poorly understood. We report that a protein at this interface, CENP-M, is structurally and evolutionarily related to small GTPases but is incapable of GTP-binding and conformational switching. We show that CENP-M is crucially required for the assembly and stability of a tetramer also comprising CENP-I, CENP-H, and CENP-K, the HIKM complex, which we extensively characterize through a combination of structural, biochemical, and cell biological approaches. A point mutant affecting the CENP-M/CENP-I interaction hampers kinetochore assembly and chromosome alignment and prevents kinetochore recruitment of the CENP-T/W complex, questioning a role of CENP-T/W as founder of an independent axis of kinetochore assembly. Our studies identify a single pathway having CENP-C as founder, and CENP-H/I/K/M and CENP-T/W as CENP-C-dependent followers.
