The deubiquitinating enzyme OTUD6A deubiquitinates the TEAD4 transcription factor, enhancing its interaction with co-activator YAP over co-repressor VGLL4. This modulation promotes TEAD4 transcriptional activity, facilitating the expression of multiple target genes controlled by the YAP-TEAD complex.

TEAD transcription factors play a central role in the Hippo signaling pathway. In this study, we focused on transcriptional enhancer factor TEF-3 (TEAD4), exploring its regulation by the deubiquitinase OTU domain-containing protein 6A (OTUD6A). We identified OTUD6A as a TEAD4-interacting deubiquitinase, positively influencing TEAD-driven transcription without altering TEAD4 stability. Structural analyses revealed specific interaction domains: the N-terminal domain of OTUD6A and the YAP-binding domain of TEAD4. Functional assays demonstrated the positive impact of OTUD6A on the transcription of YAP–TEAD target genes. Despite no impact on TEAD4 nuclear localization, OTUD6A selectively modulated nuclear interactions, enhancing YAP–TEAD4 complex formation while suppressing VGLL4 (transcription cofactor vestigial-like protein 4)–TEAD4 interaction. Critically, OTUD6A facilitated YAP–TEAD4 complex binding to target gene promoters. Our study unveils the regulatory landscape of OTUD6A on TEAD4, providing insights into diseases regulated by YAP–TEAD complexes.