Journal article
Hierarchical inactivation of a synthetic human kinetochore by a chromatin modifier
Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3JR, United Kingdom.1
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.
| Language: | English |
|---|---|
| Publisher: | The American Society for Cell Biology |
| Year: | 2009 |
| Pages: | 4194-204 |
| ISSN: | 19394586 and 10591524 |
| Types: | Journal article |
| DOI: | 10.1091/mbc.E09-06-0489 |
Autoantigens CENPA protein, human Cell Line, Tumor Centromere Centromere Protein A 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 TRIM28 protein, human Transfection Tripartite Motif-Containing Protein 28 centromere protein C
