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Characterization of induced cohesin loop extrusion trajectories in living cells [Hi-C]

ABSTRACT: Characterization of induced cohesin loop extrusion trajectories in living cells [Hi-C]

PROVIDER: PRJNA1183834 | ENA |

REPOSITORIES: ENA

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			Action	DRS
		Other
	SRR31285670_1.fastq.gz	Fastqsanger.gz
	SRR31285670_2.fastq.gz	Fastqsanger.gz
	SRR31285671_1.fastq.gz	Fastqsanger.gz
	SRR31285671_2.fastq.gz	Fastqsanger.gz

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Project description:The cohesin complex shapes chromosomes by DNA loop extrusion, but individual extrusion trajectories were so far unappreciable in vivo. Here, we developed and validated TArgeted Cohesin Loader (TACL), a system enabling the strong activation of anchored loop extrusion from dozens of defined genomic sites in living cells. Studying their individual loop extrusion trajectories revealed that extruding cohesin-STAG2 stops not only at domain boundaries but at all flanking CTCF sites, engaging them in a complex transient looping network that supports intradomain contacts. Cohesin-STAG1 cannot associate with weak CTCF binding sites and fails to similarly support intradomain interactions. NIPBL-MAU2 remains associated with cohesin when stalled at looping CTCF sites, suggesting these factors may also be required for loop stabilization. TACL induces cohesin traffic jams and illegal loops with divergent CTCF sites, demonstrating that stalled cohesin can block extruding cohesin in vivo. Genes exposed to TACL-induced loop extrusion were collectively hindered in transcription and the underlying chromatin altered its accessibility and reduced its H3K27ac marks. Thus, by enabling the study of individual loop extrusion trajectories in vivo, we could assign new functions to players, identify new looping networks and uncover an interplay between loop extrusion, gene transcription and chromatin composition.

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Project description:Characterization of induced cohesin loop extrusion trajectories in living cells

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