Induced Ectopic Kinetochore Assembly Bypasses the Requirement for CENP-A Nucleosomes
Author(s)Gascoigne, Karen E.; Takeuchi, Kozo; Suzuki, Aussie; Hori, Tetsuya; Fukagawa, Tatsuo; Cheeseman, Iain McPherson; ... Show more Show less
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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.
DepartmentMassachusetts Institute of Technology. Department of Biology; Whitehead Institute for Biomedical Research
Gascoigne, Karen E., Kozo Takeuchi, Aussie Suzuki, Tetsuya Hori, Tatsuo Fukagawa, and Iain M. Cheeseman. “Induced Ectopic Kinetochore Assembly Bypasses the Requirement for CENP-A Nucleosomes.” Cell 145, no. 3 (April 2011): 410-422. Copyright © 2011 Elsevier Inc.
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