Identifying proteins bound to native mitotic ESC chromosomes reveals chromatin repressors are important for compaction
Open Access
- 17 August 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Communications
- Vol. 11 (1), 1-15
- https://doi.org/10.1038/s41467-020-17823-z
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.Keywords
This publication has 98 references indexed in Scilit:
- Methylation-Dependent and -Independent Genomic Targeting Principles of the MBD Protein FamilyCell, 2013
- Chromatin higher-order structures and gene regulationCurrent Opinion in Genetics & Development, 2011
- The Polycomb complex PRC2 and its mark in lifeNature, 2011
- The Protein Composition of Mitotic Chromosomes Determined Using Multiclassifier Combinatorial ProteomicsCell, 2010
- Progressive Activation of CyclinB1-Cdk1 Coordinates Entry to MitosisDevelopmental Cell, 2010
- A Reconfigured Pattern of MLL Occupancy within Mitotic Chromatin Promotes Rapid Transcriptional Reactivation Following Mitotic ExitMolecular Cell, 2009
- Role of the polycomb protein EED in the propagation of repressive histone marksNature, 2009
- A model for transmission of the H3K27me3 epigenetic markNature, 2008
- Automated electron microscope tomography using robust prediction of specimen movementsJournal of Structural Biology, 2005
- Computer Visualization of Three-Dimensional Image Data Using IMODJournal of Structural Biology, 1996