Inactivation of a human kinetochore by specific targeting of chromatin modifiers

被引：215

作者：

Nakano, Megumi ^{[1
]}

Cardinale, Stefano ^{[2
]}

Noskov, Vladimir N. ^{[1
]}

Gassmann, Reto ^{[2
]}

Vagnarelli, Paola ^{[2
]}

Kandels-Lewis, Stefanie ^{[2
]}

Larionov, Vladimir ^{[1
]}

Earnshaw, William C. ^{[2
]}

Masumoto, Hiroshi ^{[1
]}

机构：

[1] NCI, Mol Pharmacol Lab, NIH, Bethesda, MD 20892 USA

[2] Univ Edinburgh, Welcome Trust Ctr Cell Biol, Edinburgh EH9 3JR, Midlothian, Scotland

来源：

DEVELOPMENTAL CELL | 2008年 / 14卷 / 04期

基金：

英国惠康基金;

关键词：

D O I：

10.1016/j.devcel.2008.02.001

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

We have used a human artificial chromosome (HAC) to manipulate the epigenetic state of chromatin within an active kinetochore. The HAC has a dimeric alpha-satellite repeat containing one natural monomer with a CENP-B binding site, and one completely artificial synthetic monomer with the CENP-B box replaced by a tetracycline operator (tetO). This HAC exhibits normal kinetochore protein composition and mitotic stability. Targeting of several tet-repressor (tetR) fusions into the centromere had no effect on kinetochore function. However, altering the chromatin state to a more open configuration with the tTA transcriptional activator or to a more closed state with the tTS transcription silencer caused missegregation and loss of the HAC. tTS binding caused the loss of CENP-A, CENP-B, CENP-C, and H3K4me2 from the centromere accompanied by an accumulation of histone H3K9me3. Our results reveal that a dynamic balance between centromeric chromatin and heterochromatin is essential for vertebrate kinetochore activity.

引用

页码：507 / 522

页数：16

共 63 条

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