abstract: Mediator is a multi-protein complex known to play a key role in eukaryotic transcriptional regulation through its interactions with transcription factors and RNA Pol II. I will present recent experimental evidence in S. cerevisiae, based on chromatin immunoprecipitation and mass spectrometry, which shows that, in addition to binding Pol II promoters, Mediator occupies chromatin domain boundaries and associates with proteins known to be involved in higher-order genome organization. These results suggest that Mediator plays a significant role in both gene regulation and establishing and maintaining 3D genome structure.