Evidence for additive and synergistic action of mammalian enhancers during cell fate determination

Enhancer activity drives cell differentiation and cell fate determination, but it remains unclear how enhancers cooperate during these processes. Here we investigate enhancer cooperation during transdifferentiation of human leukemia B-cells to macrophages. Putative enhancers are established by binding of the pioneer factor C/EBP alpha followed by chromatin opening and enhancer RNA (eRNA) synthesis from H3K4-monomethylated regions. Using eRNA synthesis as a proxy for enhancer activity, we find that most putative enhancers cooperate in an additive way to regulate transcription of assigned target genes. However, transcription from 136 target genes depends exponentially on the summed activity of its putative paired enhancers, indicating that these enhancers cooperate synergistically. The target genes are cell type-specific, suggesting that enhancer synergy can contribute to cell fate determination. Enhancer synergy appears to depend on cell type-specific transcription factors, and such interacting enhancers are not predicted from occupancy or accessibility data that are used to detect superenhancers.

Automated summary

Enhancer activity plays a crucial role in cell differentiation and cell fate determination. This study demonstrates that while most putative enhancers cooperate in an additive manner, some enhancers synergistically regulate the transcription of target genes, which are cell type-specific and can contribute to cell fate determination. Enhancer synergy appears to depend on cell type-specific transcription factors and cannot be predicted from occupancy or accessibility data used to detect superenhancers.