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

Jan H Bergmann, Mariluz Gómez Rodríguez, Nuno M C Martins, Hiroshi Kimura, David A Kelly, Hiroshi Masumoto, Vladimir Larionov, Lars E T Jansen, William C Earnshaw

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

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.
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

Keywords / Materials (for Non-textual outputs)

  • 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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