Kroeger, B.Manning, S.A.Mohan, V.Lou, J.Sun, G.Lamont, S.McCann, A.J.Francois, M.Polo, J.M.Hinde, E.Harvey, K.F.2025-10-142025-10-142025Science Advances, 2025; 11(30):eadw4974-1-eadw4974-162375-25482375-2548https://hdl.handle.net/2440/147751Over the past two decades, genetic and proteomic screens have identified the Hippo pathway as a complex signaling network that controls tissue growth and human cancer. Despite these advances, our understanding of how Hippo signaling regulates transcription is less clear. To address this, we used live microscopy to study the nuclear behavior of the major Hippo pathway transcription effectors, YAP and TEADs. We reveal that TEADs are a major determinant of YAP DNA binding and nuclear mobility, while YAP minorly influences TEADs. YAP and TEAD1 associate with DNA for longer periods in cells with intrinsically low Hippo pathway activity and upon acute Hippo pathway perturbation. TEAD1 binds the genome on a broad range of timescales, and this is extended substantially in nuclear condensates. Last, a cancer-associated YAP fusion protein exhibits substantially different biophysical behavior than either YAP or TEAD1. Thus, we reveal that Hippo signaling regulates transcription, in part, by influencing the DNA binding times of YAP and TEADs.en© 2025 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY)Hippo signaling; DNA; YAP; TEADAdaptor Proteins, Signal TransducingCell NucleusDNADNA-Binding ProteinsHippo Signaling PathwayHumansNuclear ProteinsPhosphoproteinsProtein BindingProtein Serine-Threonine KinasesSignal TransductionTEA Domain Transcription FactorsTranscription FactorsTranscription, GeneticYAP-Signaling ProteinsJournal article10.1126/sciadv.adw4974746226Polo, J.M. [0000-0002-2531-778X]