High-resolution binding data of TFIID and cofactors show promoter-specific differences <i>in vivo</i>

TFIID is instrumental in recognizing promoter sequences and initiating transcription, yet a cohesive understanding of how this complex interacts with and functions at different promoter types <i>in vivo</i> is still lacking. Here, we employed ChIP-nexus to capture high-resolution binding footprints of all <i>Drosophila</i> TFIID subunits across the genome. These footprints reveal TFIID sub-modules whose DNA contacts suggest new structural details. At different promoter types, the footprints of the TAFs are very similar, suggesting the presence of engaged TFIID across all promoters. In contrast, the binding profile of TBP is promoter-specific, enabling us to identify TATA, DPR, and TCT/housekeeping promoters <i>de novo</i>, along with their underlying core promoter elements. Notably, our data point to NC2 being specific for TBP binding at the TATA box and suggest that TATA promoters show both TAF-dependent and TAF-independent initiation <i>in vivo</i>. These data suggest a model for the increased burst size observed at TATA promoters and provide a comprehensive resource for linking structural and biochemical results to <i>in vivo</i> data.