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

Abstract:
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.
Publication status:
Published
Peer review status:
Peer reviewed

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Publisher copy:
10.1038/s41467-020-17823-z

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Role:
Author
ORCID:
0000-0002-7914-9777
More by this author
Role:
Author
ORCID:
0000-0002-6400-0945
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Role:
Author
ORCID:
0000-0002-2996-2479


Publisher:
Springer Nature
Journal:
Nature Communications More from this journal
Volume:
11
Issue:
1
Article number:
4118
Publication date:
2020-08-17
Acceptance date:
2020-07-15
DOI:
EISSN:
2041-1723
Pmid:
32807789

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