Abstract
The Hippo TEAD-transcriptional regulators YAP1 and TAZ are central for cell renewal and cancer growth; however, the specific downstream gene networks involved in their activity are not completely understood. Here we introduce TEADi, a genetically encoded inhibitor of the interaction of YAP1 and TAZ with TEAD, as a tool to characterize the transcriptional networks and biological effects regulated by TEAD transcription factors. Blockage of TEAD activity by TEADi in human keratinocytes and mouse skin leads to reduced proliferation and rapid activation of differentiation programs. Analysis of gene networks affected by TEADi and YAP1/TAZ knockdown identifies KLF4 as a central transcriptional node regulated by YAP1/TAZ-TEAD in keratinocyte differentiation. Moreover, we show that TEAD and KLF4 can regulate the activity of each other, indicating that these factors are part of a transcriptional regulatory loop. Our study establishes TEADi as a resource for studying YAP1/TAZ-TEAD dependent effects.
Highlights
The Hippo TEAD-transcriptional regulators YAP1 and TAZ are central for cell renewal and cancer growth; the specific downstream gene networks involved in their activity are not completely understood
Specific TEAD-interacting domains are present in the Hippo-family members YAP1 and TAZ, and in vestigial-like (VGLL) proteins[20,21], and it has been shown that peptides containing these domains are effective in antagonizing YAP1 activity by blocking YAP1 binding to TEAD21,22
Geneticallyencoded inhibitors give more flexibility to increases in protein size compared with soluble peptide inhibitors, allowing us to include the TEAD interaction domain of TAZ (TBD TAZ)[20] to increase inhibition towards TAZ, a bipartite nuclear localization signal (BPNLS)[23] to target the construct, and a green fluorescent protein (GFP) to track expression and localization of TEADi
Summary
The Hippo TEAD-transcriptional regulators YAP1 and TAZ are central for cell renewal and cancer growth; the specific downstream gene networks involved in their activity are not completely understood. Analysis of gene networks affected by TEADi and YAP1/TAZ knockdown identifies KLF4 as a central transcriptional node regulated by YAP1/ TAZ-TEAD in keratinocyte differentiation. YAP1 and TAZ are implicated in skin basal (BCC) and squamous (SCC) cell carcinoma formation[7,10,11] This axis has been recently listed as one of the top 10 signaling pathways altered in human cancer[12]. Studies of the role of YAP1/TAZ in skin homeostasis and cancer involve the knockout of these co-transcriptional activators, impinging on TEAD-dependent events, and potentially affecting a myriad of other transcriptional and signaling components that interact with YAP1 and TAZ16,17. To characterize the precise transcriptional events regulated downstream of YAP1/TAZ-TEAD, we introduce TEADi (TEADinhibitor), a genetically encoded fluorescently-tagged inhibitor of the interaction of YAP1 and TAZ with TEAD transcription factors. We use TEADi to identify the transcriptional networks regulated by TEAD using as a model human keratinocytes and mouse skin
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