YAP and TAZ are transcriptional co-activators of AP-1 proteins and STAT3 during breast cellular transformation
- Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, United States
- Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, United States
- Genetics Laboratory, Shenzhen Longgang District Maternity and Child Healthcare Hospital, China
Figures
Figure 1
YAP and TAZ facilitate transformation.
(A) Western blot for YAP, TAZ, and actin levels in the indicated CRISPR-mediated knockout (KO) strains and the parental cell line (CTRL). (B) Relative growth in low attachment conditions of the …
Figure 2 with 1 supplement
YAP and TAZ regulate STAT3 and JUNB activities during transformation.
(A) Western blot for the indicated proteins (STAT3-p, a form phosphorylated at Tyr705) in cells treated with the indicated siRNAs for 24 hr and then with TAM for the indicated times. (B) Normalized …
Figure 2—figure supplement 1
Gene expression in siRNA knockdowns.
RNA levels of YAP, TAZ, STAT3, and NF-κB1 in non-transformed (ETH) or transformed (TAM) cells knocked down by siRNA (i) of the indicated factors.
Figure 3 with 1 supplement
YAP and TAZ, via WW domains, directly interact with STAT3 and JUNB.
(A) Levels of the indicated proteins in the indicated fractions in non-transformed (E; ethanol) and transformed (T; tamoxifen) conditions: MEK1/2, cytoplasm marker; U1 SnRNAP70, nucleoplasm marker; …
Figure 3—figure supplement 1
Interactions of YAP and TAZ with JUNB and STAT3.
(A) Levels of the indicated proteins in the cytoplasm or nucleus of non-transformed (E) or transformed (T) cells. LDH-A is a cytoplasm marker and H3 is a nucleus marker. (B) Levels of the indicated …
Figure 4 with 1 supplement
YAP and TAZ have highly similar binding profiles, but a small subset of binding sites is unique to each factor.
(A) Correlation (r ~ 0.7) of YAP and TAZ binding signals in non-transformed (ETH) and transformed (TAM) cells. Putative YAP- and TAZ-specific sites are indicated, respectively, as green and yellow. …
Figure 4—figure supplement 1
Motif enrichment and overlap of ChIP-seq peaks.
Enrichment of the indicated motifs (different colors) in STAT3, JUNB, TEAD, and TAZ-specific (i.e., not bound by YAP) binding sites when compared with matched control DNase-hypersensitive regions in …
Figure 5 with 1 supplement
YAP and TAZ co-occupy sites with JUNB and STAT3 in transformed ER-Src cells.
(A) Heatmaps indicate ChIP-seq signals of the indicated proteins for YAP/TAZ peaks arrayed from top to bottom in decreasing YAP ChIP signal. (B) Distance between peak summits for biological …
Figure 5—figure supplement 1
Co-binding of YAP/TAZ with other factors in non-transformed ER-Src cells (ethanol grown).
(A) Heatmaps indicate ChIP-seq signals of YAP, TAZ, TEAD, JUNB, and STAT3 for YAP/TAZ peaks arrayed from top to bottom in decreasing YAP ChIP signal. (B) Distance between peak summits for biological …
Figure 6 with 3 supplements
YAP/TAZ are recruited by TEAD, AP-1, CEBP proteins, and (to a lesser extent) STAT3.
(A) Percent of binding regions for the indicated proteins that contain a given DNA sequence motif. (B) Enrichment and p-values for various motifs within YAP/TAZ target sites in transformed cells as …
Figure 6—figure supplement 1
Distribution of the distances between AP-1 and TEAD motifs at YAP/TAZ binding sites containing both motifs.
Figure 6—figure supplement 2
YAP/TAZ binding sites and their co-binding with other factors in MDA-MD-231 cells.
(A) Heatmaps indicate ChIP-seq signals of indicated proteins for YAP/TAZ peaks arrayed from top to bottom in decreasing YAP ChIP signal. (B) Correlation between YAP and TAZ binding levels, with …
Figure 6—figure supplement 3
YAP/TAZ Motif enrichment and co-binding of YAP/TAZ with other factors in MDA-MD-231 cells.
(A) Percent of binding regions for the indicated proteins that contain a given DNA sequence motif (different colors). (B) Enrichment of motifs within YAP/TAZ binding sites as compared with control …
Figure 7 with 2 supplements
Genes regulated by YAP, TAZ, JUNB, and TEAD and association of various classes of YAP/TAZ target site with differences in overall survival in triple-negative breast cancer patients.
(A) Overlap between genes with decreased or increased expression in transformed cells treated with RNAi against YAP, TAZ, STAT3, or JUNB when compared with a control RNAi. (B) Heatmap illustrates …
Figure 7—figure supplement 1
Kaplan–Meier survival curves for the indicates genes sets from (A) the complete set of YAP/TAZ target sites, (B) the subset containing only AP-1 motifs, (C) the subset containing only TEAD motifs, and (D) the subset containing both AP-1 and TEAD motifs, for the indicated cohorts of breast cancer patients (luminal A, left; luminal B, center; HER2+, right).
Figure 7—figure supplement 2
Kaplan–Meier survival curves for genes sets from (A) YAP/TAZ target sites containing exactly one AP-1 motif site, (B) YAP/TAZ target sites containing two or more AP-1 motif sites, (C) YAP/TAZ target sites containing exactly one TEAD motif site, and (D) YAP/TAZ target sites containing two or more TEAD motif sites.
Figure 8
Model for YAP/TAZ recruitment by AP-1, STAT3, and TEAD proteins.
Three different classes of YAP/TAZ target sites based on the recruiting motif are depicted. The overall complex (gray oval) includes all three DNA-binding proteins and perhaps additional proteins …
Additional files
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Supplementary file 1
Correlation between (A) biological replicates and (B) number of binding sites of the indicated proteins in ER-Src ethanol or tamoxifen; wild-type and YAP or TAZ knockout (KO) and MDA-MB-231 cells.
- https://cdn.elifesciences.org/articles/67312/elife-67312-supp1-v2.pdf
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Supplementary file 2
Enriched GO categories for YAP-specific, TAZ-specific, or shared YAP/TAZ target genes.
- https://cdn.elifesciences.org/articles/67312/elife-67312-supp2-v2.xlsx
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Supplementary file 3
Gene signatures for the indicated categories of YAP/TAZ target sites.
- https://cdn.elifesciences.org/articles/67312/elife-67312-supp3-v2.xlsx
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Supplementary file 4
Enriched GO terms with associated q-values for genes with YAP/TAZ target sites containing only AP-1, TEAD, or STAT motif but not combinations of motifs.
- https://cdn.elifesciences.org/articles/67312/elife-67312-supp4-v2.xlsx
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Supplementary file 5
siRNAs, antibodies, qPCR primers, ChIP-seq primers, and list of datasets.
- https://cdn.elifesciences.org/articles/67312/elife-67312-supp5-v2.xlsx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/67312/elife-67312-transrepform1-v2.pdf
