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| Status |
Public on Jun 01, 2020 |
| Title |
Systematic identification of factors bound to isolated metaphase ESC chromosomes reveals a role for chromatin repressors in compaction |
| Organism |
Mus musculus |
| Experiment type |
Other
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| Summary |
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.
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| Overall design |
Hi-C on isolated chromosomes
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| Contributor(s) |
Djeghloul D, Guo Y, Merkenschlager M, Fisher AG |
| Citation(s) |
32807789 |
| |
| Submission date |
Aug 30, 2019 |
| Last update date |
Aug 31, 2020 |
| Contact name |
Ya Guo |
| Organization name |
MRC London Institute of Medical Sciences
|
| Department |
Epigenetics section
|
| Lab |
Lymphocyte Development
|
| Street address |
Faculty of Medicine, Imperial College, Hammersmith Hospital Campus
|
| City |
London |
| ZIP/Postal code |
W12 0NN |
| Country |
United Kingdom |
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| Platforms (1) |
| GPL17021 |
Illumina HiSeq 2500 (Mus musculus) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA563054 |
| SRA |
SRP219921 |
