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Epigenetic engineering shows H3K4me2 is required for HJURP targeting and CENP-A assembly on a synthetic human kinetochore

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Original languageEnglish
Pages (from-to)328-340
Number of pages13
JournalEMBO Journal
Volume30
Issue number2
Early online date14 Dec 2010
DOIs
Publication statusPublished - Jan 2011

Abstract

Kinetochores assemble on distinct 'centrochromatin' containing the histone H3 variant CENP-A and interspersed nucleosomes dimethylated on H3K4 (H3K4me2). Little is known about how the chromatin environment at active centromeres governs centromeric structure and function. Here, we report that centrochromatin resembles K4-K36 domains found in the body of some actively transcribed housekeeping genes. By tethering the lysine-specific demethylase 1 (LSD1), we specifically depleted H3K4me2, a modification thought to have a role in transcriptional memory, from the kinetochore of a synthetic human artificial chromosome (HAC). H3K4me2 depletion caused kinetochores to suffer a rapid loss of transcription of the underlying α-satellite DNA and to no longer efficiently recruit HJURP, the CENP-A chaperone. Kinetochores depleted of H3K4me2 remained functional in the short term, but were defective in incorporation of CENP-A, and were gradually inactivated. Our data provide a functional link between the centromeric chromatin, α-satellite transcription, maintenance of CENP-A levels and kinetochore stability.

    Research areas

  • Autoantigens, Centromere, Chromatin, Chromatin Immunoprecipitation, Chromosomal Proteins, Non-Histone, Chromosomes, Artificial, Human, DNA Primers, DNA-Binding Proteins, Epigenesis, Genetic, Genetic Engineering, Histones, Humans, Kinetochores, Nucleosomes, Reverse Transcriptase Polymerase Chain Reaction

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