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

Authors :
Djeghloul D
Patel B
Kramer H
Dimond A
Whilding C
Brown K
Kohler AC
Feytout A
Veland N
Elliott J
Bharat TAM
Tarafder AK
Löwe J
Ng BL
Guo Y
Guy J
Huseyin MK
Klose RJ
Merkenschlager M
Fisher AG
Source :
Nature communications [Nat Commun] 2020 Aug 17; Vol. 11 (1), pp. 4118. Date of Electronic Publication: 2020 Aug 17.
Publication Year :
2020

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.

Details

Language :
English
ISSN :
2041-1723
Volume :
11
Issue :
1
Database :
MEDLINE
Journal :
Nature communications
Publication Type :
Academic Journal
Accession number :
32807789
Full Text :
https://doi.org/10.1038/s41467-020-17823-z