Hotspot mutations in the structured ENL YEATS domain link aberrant transcriptional condensates and cancer

This data submission contains the raw microscopy images and western blot images for our manuscript, which describes the following: Growing evidence suggests prevalence of transcriptional condensates on chromatin, yet their mechanisms of formation and functional significance in disease remain unclear. In human cancer, a series of mutations in the histone acetylation reader ENL create gain-of-function mutants with increased transcriptional activation ability. Here we show that these mutations, clustered in ENL’s structured acetyl-reading YEATS domain, trigger aberrant condensates at native genomic targets through multivalent homotypic and heterotypic interactions. Mechanistically, mutation-induced structural changes in the YEATS domain, ENL’s two disordered regions of opposing charges, and the incorporation of extrinsic elongation factors are all required for ENL condensate formation. Extensive mutagenesis establishes condensate formation as a driver of oncogenic gene activation. Furthermore, expression of ENL mutants beyond the endogenous level leads to non-functional condensates. Our findings provide new mechanistic and functional insights into cancer-associated condensates and support condensate dysregulation as an oncogenic mechanism.