TY - JOUR
T1 - The pseudo GTPase CENP-M drives human kinetochore assembly
AU - Basilico, Federica
AU - Maffini, Stefano
AU - Weir, John R.
AU - Prumbaum, Daniel
AU - Rojas, Ana M.
AU - Zimniak, Tomasz
AU - De Antoni, Anna
AU - Jeganathan, Sadasivam
AU - Voss, Beate
AU - Van Gerwen, Suzan
AU - Krenn, Veronica
AU - Massimiliano, Lucia
AU - Valencia, Alfonso
AU - Vetter, Ingrid R.
AU - Herzog, Franz
AU - Raunser, Stefan
AU - Pasqualato, Sebastiano
AU - Musacchio, Andrea
N1 - Copyright © 2014, Basilico et al.
PY - 2014/7/8
Y1 - 2014/7/8
N2 - 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.
AB - 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.
KW - Amino Acid Sequence
KW - Cell Cycle Proteins
KW - Chromosomal Proteins, Non-Histone/chemistry
KW - Crystallography, X-Ray
KW - GTP Phosphohydrolases/chemistry
KW - HeLa Cells
KW - Humans
KW - Kinetochores/chemistry
KW - Models, Biological
KW - Models, Molecular
KW - Molecular Sequence Data
KW - Multiprotein Complexes/chemistry
KW - Mutation
KW - Nuclear Proteins/chemistry
KW - Protein Folding
KW - Protein Stability
KW - Protein Structure, Quaternary
KW - Protein Subunits
KW - RNA, Small Interfering/genetics
KW - Sequence Homology, Amino Acid
UR - http://www.scopus.com/inward/record.url?scp=84904052265&partnerID=8YFLogxK
U2 - 10.7554/eLife.02978.001
DO - 10.7554/eLife.02978.001
M3 - Article
C2 - 25006165
AN - SCOPUS:84904052265
SN - 2050-084X
VL - 2014
SP - e02978
JO - eLife
JF - eLife
IS - 3
M1 - e02978
ER -
