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| Status |
Public on Feb 27, 2022 |
| Title |
Cohesin and CTCF control chromosomal contact dynamics in living cells [4C-seq] |
| Organism |
Mus musculus |
| Experiment type |
Other
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| Summary |
Mammalian chromosomes are partitioned into topologically associating domains (TADs) by the loop-extrusion activity of cohesin that is blocked at specific DNA sites bound by CTCF. Chromosome structure inside TADs is highly variable in single cells, yet little is known about its temporal dynamics, how it influences the rates and durations of chromosomal contacts, and how it depends on CTCF and cohesin. To address these questions we combine two quantitative live-cell imaging strategies that minimize locus-specific confounding effects. We show that loop extrusion by cohesin globally reduces the mobility of the chromatin fiber in living cells, while also increasing the rates of formation and durations of contacts between sequences inside the same TAD. Quantitative analysis of high-resolution microscopy data reveals that contacts assemble and disassemble frequently in the course of the cell cycle, and become substantially more frequent and longer in the presence of convergent CTCF sites. Comparison with polymer modeling additionally reveals that cohesin-mediated CTCF loops last around 10 minutes on average. Our data support the notion that chromosome structure within TADs is highly dynamic and provide a quantitative framework for understanding the principles that link chromosome structure to biological function.
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| Overall design |
We performed 4C experiments in cell lines carrying random 3xCTCF-TetO insertions in the genome, to show that new interactions were formed between the 3xCTCF and endogenous CTCF sites and to show that Cre recombination and/or Rad21 depletion led to loss of these interactions.
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| Contributor(s) |
Mach P, Kos P, Zhan Y, Cramard J, Gaudin S, Tünnermann J, Eglinger J, Zuin J, Kryzhanovska M, Smallwood S, Gelman L, Marchi E, Nora EP, Tiana G, Giorgetti L |
| Citation(s) |
36471076 |
| |
| Submission date |
Feb 22, 2022 |
| Last update date |
Jun 01, 2024 |
| Contact name |
Luca Giorgetti |
| Organization name |
Friedrich Miescher Institute for Biomedical Research
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| Street address |
Fabrikstrasse 24
|
| City |
Basel |
| ZIP/Postal code |
4056 |
| Country |
Switzerland |
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| Platforms (1) |
| GPL17021 |
Illumina HiSeq 2500 (Mus musculus) |
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| Samples (6)
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| This SubSeries is part of SuperSeries: |
| GSE197238 |
Cohesin and CTCF control chromosomal contact dynamics in living cells |
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| Relations |
| BioProject |
PRJNA809391 |
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