Centromere transcription allows CENP-A to transit from chromatin association to stable incorporation

Georg O.M. Bobkov, Nick Gilbert, Patrick Heun

Research output: Contribution to journalArticlepeer-review

Abstract

Centromeres are essential for chromosome segregation and are specified epigenetically by the presence of the histone H3 variant CENP-A. In flies and humans, replenishment of the centromeric mark is uncoupled from DNA replication and requires the removal of H3 "placeholder" nucleosomes. Although transcription at centromeres has been previously linked to the loading of new CENP-A, the underlying molecular mechanism remains poorly understood. Here, we used Drosophila melanogaster tissue culture cells to show that centromeric presence of actively transcribing RNA polymerase II temporally coincides with de novo deposition of dCENP-A. Using a newly developed dCENP-A loading system that is independent of acute transcription, we found that short inhibition of transcription impaired dCENP-A incorporation into chromatin. Interestingly, initial targeting of dCENP-A to centromeres was unaffected, revealing two stability states of newly loaded dCENP-A: a salt-sensitive association with the centromere and a salt-resistant chromatin-incorporated form. This suggests that transcription-mediated chromatin remodeling is required for the transition of dCENP-A to fully incorporated nucleosomes at the centromere.
Original languageEnglish
Pages (from-to)1957–1972
Number of pages16
JournalJournal of Cell Biology
Volume217
Issue number6
Early online date6 Apr 2018
DOIs
Publication statusPublished - 1 Jun 2018

Keywords

  • chromatin or epigenetics
  • cell cycle and division

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