Extrusion fountains are hallmarks of chromosome organization emerging upon zygotic genome activation

TL;DR

Chromosome folding in early embryos starts with 'fountains' at enhancers, linked to cohesin loading and transcription factor activity.

Contribution

Identifies 'fountains' as a new hallmark of chromosome organization emerging during zygotic genome activation.

Findings

Fountains form early at enhancers and depend on cohesin accumulation.

Pioneer transcription factors are essential for fountain formation.

Fountains reappear after mitosis and are observed in mouse cells.

Abstract

The initiation of gene expression during development, known as zygotic genome activation (ZGA), is accompanied by massive changes in chromosome organization. However, the earliest events of chromosome folding and their functional roles remain unclear. Using Hi-C on zebrafish embryos, we discovered that chromosome folding begins early in development with the formation of fountains, distinct elements of chromosome organization. Emerging preferentially at enhancers, fountains show an initial accumulation of cohesin, which later redistributes to CTCF sites at TAD borders. Knockouts of pioneer transcription factors driving ZGA enhancers cause a specific loss of fountains, establishing a causal link between enhancer activation and fountain formation. Polymer simulations demonstrate that fountains may arise as sites of facilitated cohesin loading, requiring two-sided but desynchronized loop extrusion, potentially caused by cohesin collisions with obstacles or internal switching. Moreover, we detected cohesin-dependent fountain patterns at enhancers in mouse cells and found them reemerging with cohesin loading after mitosis. Altogether, fountains represent enhancer-specific elements of chromosome organization and suggest that chromosome folding during development and after cell division starts with facilitated cohesin loading. Observations in multiple systems further support facilitated loading at enhancers as a widespread phenomenon.

Chromosome folding in early zebrafish development starts with formation of fountains. These patterns, also prominent in other systems, form at active enhancers, depend on transcription factors, and reflect preferential loading of loop extruding cohesin at enhancers.

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