Endogenous YAP/TAZ partitioning in TEAD condensates orchestrates the Hippo response

YAP/TAZ are transcriptional co-activators that pair with transcription factor TEA/ATTS domains (TEADs) for modulating the Hippo pathway. Previous works propose the potential role of YAP/TAZ phase separation for transcriptional activation, yet the biomolecular basis of endogenous YAP/TAZ-TEAD condensates remains unclear. Here, we dissect their endogenous morphology, revealing that YAP/TAZ are client proteins recruited to TEAD condensates in various human cell lines. TEAD proteins have robust intrinsic potential to undergo phase separation, and YAP/TAZ condensates disappear immediately after losing their interaction with TEADs. Moreover, TEAD condensates serve as central organizing hubs to dynamically concentrate active YAP and other markers of transcriptional activation. Based on this, we revisited a series of recently characterized TEAD inhibitors and identified that VGLL4 represents a critical regulator assisting TEAD central pocket inhibitors. Altogether, we demonstrate a fundamental role of TEAD condensates in spatially regulating YAP/TAZ signaling, underscoring their significance in further deciphering TEAD biology and applications in TEAD-targeted therapy.