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Cell. 2011 Apr 29;145(3):410-22. doi: 10.1016/j.cell.2011.03.031.

Induced ectopic kinetochore assembly bypasses the requirement for CENP-A nucleosomes.

Author information

1
Whitehead Institute for Biomedical Research, Department of Biology, Nine Cambridge Center, MA 02142, USA.

Abstract

Accurate chromosome segregation requires assembly of the multiprotein kinetochore complex at centromeres. Although prior work identified the centromeric histone H3-variant CENP-A as the important upstream factor necessary for centromere specification, in human cells CENP-A is not sufficient for kinetochore assembly. Here, we demonstrate that two constitutive DNA-binding kinetochore components, CENP-C and CENP-T, function to direct kinetochore formation. Replacing the DNA-binding regions of CENP-C and CENP-T with alternate chromosome-targeting domains recruits these proteins to ectopic loci, resulting in CENP-A-independent kinetochore assembly. These ectopic kinetochore-like foci are functional based on the stoichiometric assembly of multiple kinetochore components, including the microtubule-binding KMN network, the presence of microtubule attachments, the microtubule-sensitive recruitment of the spindle checkpoint protein Mad2, and the segregation behavior of foci-containing chromosomes. We additionally find that CENP-T phosphorylation regulates the mitotic assembly of both endogenous and ectopic kinetochores. Thus, CENP-C and CENP-T form a critical regulated platform for vertebrate kinetochore assembly.

PMID:
21529714
PMCID:
PMC3085131
DOI:
10.1016/j.cell.2011.03.031
[Indexed for MEDLINE]
Free PMC Article

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