RT Journal Article
SR Electronic
T1 Systematic identification of factors bound to isolated metaphase ESC chromosomes reveals a role for chromatin repressors in compaction
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 750067
DO 10.1101/750067
A1 Dounia Djeghloul
A1 Anne-Céline Kohler
A1 Bhavik Patel
A1 Holger Kramer
A1 Nicolas Veland
A1 Chad Whilding
A1 Andrew Dimond
A1 James Elliott
A1 Amelie Feytout
A1 Tanmay A.M. Bharat
A1 Abul K. Tarafder
A1 Jan Löwe
A1 Bee L. Ng
A1 Ya Guo
A1 Karen Brown
A1 Jacky Guy
A1 Matthias Merkenschlager
A1 Amanda G. Fisher
YR 2019
UL http://biorxiv.org/content/early/2019/08/28/750067.abstract
AB Epigenetic information is transmitted from mother to daughter cells through mitosis. To identify trans-acting factors and cis-acting elements that might be important for conveying epigenetic memory through cell division, we isolated native (unfixed) chromosomes from metaphase-arrested cells using flow cytometry and performed LC-MS/MS to determine the repertoire of chromosome-bound proteins. Quantitative proteomic comparisons between metaphase-arrested cell lysates and chromosome-sorted samples revealed 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. We showed that deletion of PRC2, DNMT1/3a/3b or Mecp2 provoked an increase in the size of individual mitotic chromosomes consistent with de-condensation, as did experimental cleavage of cohesin complexes. These data provide a comprehensive inventory of chromosome-bound factors in pluripotent stem cells at mitosis and reveal an unexpected role for chromatin repressor complexes in preserving mitotic chromosome compaction.