@article{604adc0378394df192e0161f02cf0d9f,
title = "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.",
author = "Dounia Djeghloul and Bhavik Patel and Holger Kramer and Andrew Dimond and Chad Whilding and Karen Brown and Kohler, {Anne C{\'e}line} and Amelie Feytout and Nicolas Veland and James Elliott and Bharat, {Tanmay A.M.} and Tarafder, {Abul K.} and Jan L{\"o}we and Ng, {Bee L.} and Ya Guo and Jacky Guy and Huseyin, {Miles K.} and Klose, {Robert J.} and Matthias Merkenschlager and Fisher, {Amanda G.}",
note = "Funding Information: We would like to thank Richard Henderson for his encouragement to begin these studies, C. Tyler-Smith for enabling chromosome sorting, T. Adejumo, R. Maggio, F. Pereira, P. Chana, H. Pallikonda and N. Festuccia for expertise and providing reagents, A. Lisini for reading the manuscript and advice. We thank the LMS/NIHR Imperial Biomedical Research Centre Flow Cytometry Facility, as well as the LMS Genomics and LMS Bioinformatics facilities for support. This work was funded by core support from the Medical Research Council UK to the London Institute of Medical Sciences. R.J.K. was supported by the Wellcome Trust (209400/Z/17/Z) and the European Research Council (681440), and M.K.H. was supported by the Wellcome Trust (109102/Z/15/Z). Publisher Copyright: {\textcopyright} 2020, The Author(s).",
year = "2020",
month = aug,
day = "17",
doi = "10.1038/s41467-020-17823-z",
language = "English",
volume = "11",
journal = "Nature Communications",
issn = "2041-1723",
publisher = "Nature Publishing Group",
number = "1",
}