Hormone-induced enhancer assembly requires an optimal level of hormone receptor multivalent interactions

Transcription factors (TFs) activate enhancers to drive cell-specific gene programs in response to signals, but our understanding of enhancer assembly during signaling events is incomplete. Here, we show that androgen receptor (AR) forms condensates through multivalent interactions mediated by its N-terminal intrinsically disordered region (IDR) to orchestrate enhancer assembly in response to androgen signaling. AR IDR can be substituted by IDRs from selective proteins for AR condensation capacity and its function on enhancers. Expansion of the poly(Q) track within AR IDR results in a higher AR condensation propensity as measured by multiple methods, including live-cell single-molecule microscopy. Either weakening or strengthening AR condensation propensity impairs its heterotypic multivalent interactions with other enhancer components and diminishes its transcriptional activity. Our work reveals the requirement of an optimal level of AR condensation in mediating enhancer assembly and suggests that alteration of the fine-tuned multivalent IDR-IDR interactions might underlie AR-related human pathologies. [Display omitted] •Androgen-induced AR condensation depends on its disordered NTD and structured LBD•AR IDR can be substituted by selective IDRs for condensation and transactivation•Poly(Q) expansion in AR IDR enhances condensation but impairs enhancer assembly•An optimal level of AR multivalent interactions ensures AR enhancer regulation It is now recognized that multivalent interactions of transcriptional proteins can mediate protein recruitment on enhancers. However, the precise regulation of these interactions remains unclear. By studying androgen-responsive enhancers, Chen et al. show that an optimal level of multivalent interactions of androgen receptor is required for enhancer assembly and activation.