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Hierarchical inactivation of a synthetic human kinetochore by a chromatin modifier

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Original languageEnglish
Pages (from-to)4194-204
Number of pages11
JournalMolecular Biology of the Cell
Volume20
Issue number19
DOIs
Publication statusPublished - 2009

Abstract

We previously used a human artificial chromosome (HAC) with a synthetic kinetochore that could be targeted with chromatin modifiers fused to tetracycline repressor to show that targeting of the transcriptional repressor tTS within kinetochore chromatin disrupts kinetochore structure and function. Here we show that the transcriptional corepressor KAP1, a downstream effector of the tTS, can also inactivate the kinetochore. The disruption of kinetochore structure by KAP1 subdomains does not simply result from loss of centromeric CENP-A nucleosomes. Instead it reflects a hierarchical disruption of the outer kinetochore, with CENP-C levels falling before CENP-A levels and, in certain instances, CENP-H being lost more readily than CENP-C. These results suggest that this novel approach to kinetochore dissection may reveal new patterns of protein interactions within the kinetochore.

    Research areas

  • Autoantigens, Cell Line, Tumor, Centromere, Chromatin, Chromatin Immunoprecipitation, Chromosomal Proteins, Non-Histone, Chromosomes, Artificial, Human, HeLa Cells, Humans, Hybrid Cells, In Situ Hybridization, Fluorescence, Kinetochores, Luminescent Proteins, Microtubule-Associated Proteins, Nucleosomes, Repressor Proteins, Transfection

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