Identifying proteins bound to native mitotic ESC chromosomes reveals chromatin repressors are important for compaction

Dounia Djeghloul, Bhavik Patel, Holger Kramer, Andrew Dimond, Chad Whilding, Karen Brown, Anne Céline Kohler, Amelie Feytout, Nicolas Veland, James Elliott, Tanmay A.M. Bharat, Abul K. Tarafder, Jan Löwe, Bee L. Ng, Ya Guo, Jacky Guy, Miles K. Huseyin, Robert J. Klose, Matthias Merkenschlager, Amanda G. Fisher*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Epigenetic information is transmitted from mother to daughter cells through mitosis. Here, to identify factors that might play a role in conveying epigenetic memory through cell division, we report on the isolation of unfixed, native chromosomes from metaphase-arrested cells using flow cytometry and perform LC-MS/MS to identify chromosome-bound proteins. A quantitative proteomic comparison between metaphase-arrested cell lysates and chromosome-sorted samples reveals a cohort of proteins that were significantly enriched on mitotic ESC chromosomes. These include pluripotency-associated transcription factors, repressive chromatin-modifiers such as PRC2 and DNA methyl-transferases, and proteins governing chromosome architecture. Deletion of PRC2, Dnmt1/3a/3b or Mecp2 in ESCs leads to an increase in the size of individual mitotic chromosomes, consistent with de-condensation. Similar results were obtained by the experimental cleavage of cohesin. Thus, we identify chromosome-bound factors in pluripotent stem cells during mitosis and reveal that PRC2, DNA methylation and Mecp2 are required to maintain chromosome compaction.

Original languageEnglish
Article number4118
Number of pages15
JournalNature Communications
Volume11
Issue number1
DOIs
Publication statusPublished - 17 Aug 2020

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