Inactivation of a Human Kinetochore by Specific Targeting of Chromatin Modifiers

Overview

Journal Dev Cell

Publisher Cell Press

Specialties Cell Biology
Reproductive Medicine

Date 2008 Apr 16

PMID 18410728

Citations 170

Authors

Megumi Nakano

Stefano Cardinale

Vladimir N Noskov

Reto Gassmann

Paola Vagnarelli

Stefanie Kandels-Lewis

Vladimir Larionov

William C Earnshaw

Hiroshi Masumoto

Affiliations

Soon will be listed here.

Abstract

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.

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