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, 24 (3), 235-40

Structural Insights Into the YAP and TEAD Complex

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Structural Insights Into the YAP and TEAD Complex

Ze Li et al. Genes Dev.

Abstract

The Yes-associated protein (YAP) transcriptional coactivator is a key regulator of organ size and a candidate human oncogene inhibited by the Hippo tumor suppressor pathway. The TEAD family of transcription factors binds directly to and mediates YAP-induced gene expression. Here we report the three-dimensional structure of the YAP (residues 50-171)-TEAD1 (residues 194-411) complex, in which YAP wraps around the globular structure of TEAD1 and forms extensive interactions via three highly conserved interfaces. Interface 3, including YAP residues 86-100, is most critical for complex formation. Our study reveals the biochemical nature of the YAP-TEAD interaction, and provides a basis for pharmacological intervention of YAP-TEAD hyperactivation in human diseases.

Figures

Figure 1.
Figure 1.
Overall structure of the YAP–TEAD complex and their sequence conservation. (A) Overall structure of the human YAP–TEAD complex shown as a ribbon representation. TEAD is shown in light blue, and different YAP elements are shown in yellow, green, cyan, and red. Secondary structural elements are labeled, and two different views of the complex structure are shown. (B) Sequence alignment of TEAD and YAP across isoforms and species. TEAD, YAP, and TAZ from indicated species are included. Identical residues are highlighted with a purple background, and highly conserved residues are highlighted with a pink background. Secondary structural elements are colored as in A and are indicated above the sequences. Residues that are involved in interactions on interfaces 1, 2, and 3 (Fig. 2A) are indicated by yellow squares, green dots, and red triangles, respectively.
Figure 2.
Figure 2.
Binding interfaces between YAP and TEAD. (A) Overall interaction between YAP and TEAD. The color scheme is the same as in Figure 1A. Major binding interfaces between YAP and TEAD are highlighted as interface 1, interface 2, and interface 3. (BD) Detailed interactions between YAP and TEAD in interfaces 1, 2, and 3. Side chains involved in interaction are shown and colored as in Figure 1A. Residue numbers for YAP and TEAD are labeled in black and blue, respectively. Hydrogen bonds are represented by magenta dotted lines. (E) Sequence conservation projected on the surface of TEAD. TEAD is shown as a surface model and is colored according to the conservation scores. YAP is shown as a ribbon and is colored as in Figure 1A.
Figure 3.
Figure 3.
Effect of YAP mutations on YAP–TEAD-binding affinity. (A) In vitro pull-down assay. His-tagged YAP (50–171) and mutants were tested in a GST pull-down assay using GST-TEAD (194–411) immobilized on glutathione resin. Pull-down products were analyzed by Tricine-SDS-PAGE and Coomassie blue staining. (B) In vitro GST-TEAD pull-down of different YAP fragments. Three YAP fragments, as indicated, were expressed and purified as SUMO fusions. In vitro pull-down was performed as described in A and was analyzed by SDS-PAGE and Coomassie blue staining. (C) CTGF reporter assay. Indicated plasmids were cotransfected with a CTGF reporter and a CMV–β-gal construct into 293T cells. Luciferase activity was measured and normalized to β-galactosidase activity. YAP and TEAD expression levels were determined by Western blot with anti-Flag and anti-Myc antibodies, respectively. (D) Coimmunoprecipitation. Flag-YAP wild type or mutants were cotransfected with Myc-TEAD1 wild type into HEK293 cells. Flag-YAP was immunoprecipitated, and coimmunoprecipitated TEAD1 was determined by anti-Myc Western blot.
Figure 4.
Figure 4.
Effect of TEAD mutations on YAP–TEAD-binding affinity. (A) In vitro pull-down assay. Experiments were similar to Figure 3A except His-tagged YAP (50–171) wild-type protein and GST-TEAD1 (194–411) mutants were used. The GST fragment present in the samples of GST-TEAD1 protein preparation is indicated. (B) CTGF reporter assay. Experiments were performed as in Figure 3C with TEAD1 mutants and wide-type YAP. (C) Coimmunoprecipitation. Experiments were performed as in Figure 3D with TEAD1 mutants and wide-type YAP.

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