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Biology subjects

Nora, E.

Publications and source records attributed to Nora, E..

3 recordsLinked to original sources

Cohesin bridging as a physical principle of enhancer-promoter communication

Central to genome function, enhancers are non-coding sequences that can control transcription from promoters hundreds of kilobases away. Yet the physical basis of this long-range communication remains unclear. A prevalent view is that enhancers activate promoters when the two elements come into spatial proximity through the 3D folding of chromatin. However, activation by spatial proximity alone has struggled to explain several core features of enhancer function. Here, we propose that the molecular motor cohesin transmits long-range enhancer action by forming bridges between enhancers and promoters during loop extrusion. In this view, rare and transient bridges carry regulatory communication, rather than mere spatial proximity. We develop a quantitative model that predicts transcriptional output from cohesin-bridging dynamics and validate it by engineering cells in which strategically positioned CTCF sites rewire loop extrusion trajectories. The model explains how enhancer action scales with genomic distance, and how it can be either facilitated or insulated by CTCF sites across two orders of magnitude-behaviors incompatible with proximity-based models. Finally, our framework reveals that CTCF sites can block enhancers bidirectionally, by either blocking or releasing cohesin loops, resolving longstanding paradoxes between their effects on transcriptional regulation and genome folding. Together, our results establish cohesin bridging as a mode of enhancer-promoter communication that can be modulated by genomic context to achieve selective and tunable transcriptional control over long genomic distances.

molecular biology↗

Beyond TADs: microscopy reveals 2 axes of chromosome organization by CTCF and cohesin

While population level analyses reveal significant roles for CTCF and cohesin in mammalian genome organization, their contribution to chromatin structure and gene regulation at the single-cell level remain incompletely understood 1-4. Here, we use a super-resolution microscopy approach, Optical Reconstruction of Chromatin Architecture (ORCA) 5 to measure the effects of removal of CTCF or cohesin on genome folding across genomic scales. In untreated embryonic stem cells, we observe intricate, frequently stacked loops of chromatin which are largely dissolved upon cohesin removal. The loops compact chromatin at the < 3 Mb scale, increasing proximity between sequences not only within but also between TADs. We find multi-way contacts among loop anchors, preferentially at TAD borders, and these hubs largely dissolve upon CTCF degradation. CTCF-hubs bridge intervening TAD boundaries while keeping border distal regions from neighboring TADs apart outside the hub. Cohesin dependent loops at the < 3 Mb scale impede mixing at larger chromosomal scales through steric effects of loop stacking, dramatically reducing genomic cross-talk. Disruption of this ordered chromosomal structure led to increased cell-cell variability in gene expression, exceeding changes to average expression. Together our data revise the TAD-centric understanding of CTCF and cohesin, and provide a multi-scale, structural picture of how they organize the genome on the single-cell level through distinct contributions to loop stacking.

biophysics↗

CTCF is a Barrier for Totipotent-like Reprogramming

Totipotent cells have the ability of generating embryonic and extra-embryonic tissues1,2. Interestingly, a rare population of cells with totipotent-like potential was identified within ESC cultures3. These cells, known as 2 cell (2C)-like cells, arise from ESC and display similar features to those found in the totipotent 2 cell embryo2-4. However, the molecular determinants of 2C-like conversion have not been completely elucidated. Here, we show that CTCF is a barrier for 2C-like reprogramming. Indeed, forced conversion to a 2C-like state by DUX expression was associated with DNA damage at a subset of CTCF binding sites. Endogenous or DUX-induced 2C-like ESC showed decreased CTCF enrichment at known binding sites, suggesting that acquisition of a totipotent-like state is associated with a highly dynamic chromatin architecture. Accordingly, depletion of CTCF in ESC efficiently promoted spontaneous and asynchronous conversion to a totipotent-like state. This phenotypic reprogramming was reversible upon restoration of CTCF levels. Furthermore, we showed that transcriptional activation of the ZSCAN4 cluster was necessary for successful 2C-like reprogramming. In summary, we revealed the intimate relation between CTCF and totipotent-like reprogramming.

developmental biology↗