Asymmetric Notch signaling promotes divergent fates in select cells throughout metazoan development. In the receiving cell, signaling results in cleavage of the Notch intracellular domain and its import into the nucleus, where it binds Suppressor of Hairless [Su(H)] to promote gene expression in conjunction with contextual cues in the surrounding DNA sequence. To investigate the nature of this contextual logic, we identify 1344 Su(H)-site containing regulatory belts that are conserved across the Drosophila genus. Each Su(H)-type regulatory belt (SUH-RB) is a 0.6–1.0 kb chain of conservation peaks consistent with a transcriptional enhancer or core promoter. These regulatory belts contain one or more canonical binding sites for Su(H) along with ~15–30 other binding sites. SUH-RBs are densely clustered in certain chromosomal regions such as the E(spl)-complex, the Wnt gene complex, and genes encoding Notch receptor ligands (Delta and Serrate). SUH-RBs overlap most known Su(H)/Notch-target enhancers and others, including non-embryonic enhancers that are not identified by embryonic ChIP-seq peaks. Thus, SUH-RBs overcome the stage-specific nature of embryonic ChIP-seq peaks and suggest a pervasive role for contextual tissue-specific pioneer and/or enhancer-licensing factors. SUH-RBs also delineate false positive ChIP-seq peaks, which do not overlap SUH-RBs, are missing even the weakest Su(H)-binding sequences, and have the shortest ChIP peak widths. Last, we characterize several novel enhancers including Su(H)-dependent enhancers at Notch and Delta, intestinal enhancers at A2bp1 and hedgehog, and distinct enhancers at roughest, E2f1, and escargot.

Genomic dimensions of Su(H)-targeted regulatory belts in Drosophila