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Identifying proteins bound to native mitotic ESC chromosomes reveals chromatin repressors are important for compaction.
- 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.
- Subjects :
- Animals
DNA (Cytosine-5-)-Methyltransferase 1 metabolism
DNA (Cytosine-5-)-Methyltransferases metabolism
DNA Methylation genetics
DNA Methylation physiology
DNA Methyltransferase 3A
Fluorescent Antibody Technique
Methyl-CpG-Binding Protein 2 metabolism
Mice
Proteomics
DNA Methyltransferase 3B
Chromatin metabolism
Chromosomes metabolism
Embryonic Stem Cells metabolism
Transcription Factors metabolism
Subjects
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