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.

Basilico, F., Maffini, S., Weir, J., Prumbaum, D., Rojas, A., Zimniak, T., et al. (2014). The pseudo GTPase CENP-M drives human kinetochore assembly. ELIFE, 2014(3), 1-28 [10.7554/eLife.02978].

The pseudo GTPase CENP-M drives human kinetochore assembly

Krenn, Veronica;
2014

Abstract

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.
Articolo in rivista - Articolo scientifico
centromeres; kinetochores; mitosis; Amino Acid Sequence; Cell Cycle Proteins; Chromosomal Proteins, Non-Histone; Crystallography, X-Ray; GTP Phosphohydrolases; HeLa Cells; Humans; Kinetochores; Models, Biological; Models, Molecular; Molecular Sequence Data; Multiprotein Complexes; Mutation; Nuclear Proteins; Protein Folding; Protein Stability; Protein Structure, Quaternary; Protein Subunits; RNA, Small Interfering; Sequence Homology, Amino Acid
English
2014
2014
3
1
28
e02978
open
Basilico, F., Maffini, S., Weir, J., Prumbaum, D., Rojas, A., Zimniak, T., et al. (2014). The pseudo GTPase CENP-M drives human kinetochore assembly. ELIFE, 2014(3), 1-28 [10.7554/eLife.02978].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/397677
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