Epigenetic Cell Fate in Candida albicans is Controlled by Transcription Factor Condensates Acting at Super-Enhancer-Like Elements

Cell identity in eukaryotes is controlled by transcriptional regulatory networks (TRNs) that define cell type-specific gene expression. In the opportunistic fungal pathogen Candida albicans, TRNs regulate epigenetic switching between two alternative cell states, ‘white’ and ‘opaque’, that exhibit distinct host interactions. Here, we reveal that the transcription factors (TFs) regulating cell identity contain prion-like domains (PrLDs) that enable liquid-liquid demixing and the formation of phase-separated condensates. Multiple white-opaque TFs can co-assemble into complex condensates as observed on single DNA molecules. Moreover, heterotypic interactions between PrLDs supports the assembly of multifactorial condensates at a synthetic locus within live eukaryotic cells. Mutation of the Wor1 PrLD revealed that substitution of acidic residues abolished its ability to phase separate and to co-recruit other TFs in live cells, as well as its function in C. albicans cell fate determination. Together, these studies reveal that PrLDs support the assembly of TF complexes that control fungal cell identity and highlight parallels with the ‘super-enhancers’ that regulate mammalian cell fate.