Identification of Wnt‐regulated genes that are repressed by, or independent of, β‐catenin

Wnt signaling regulates metazoan development and homeostasis, in part by β-catenin-dependent activation and repression of a large number of genes. However, Wnt signaling also regulates genes independent of β-catenin, genes that are less well-characterized. In this study, using a pan-Wnt inhibitor, we performed a comprehensive transcriptome analysis in a Wnt-addicted orthotopic cancer model to delineate the β-catenin-dependent and independent arms of Wnt signaling. We find that while a large percentage of Wnt-regulated genes are regulated by β-catenin, 10% of these genes are regulated independent of β-catenin. Interestingly, a large proportion of these β-catenin-independent genes are Wnt-repressed. Among the β-catenin-dependent genes, more than half are repressed by β-catenin. We used this dataset to investigate the mechanisms by which Wnt/β-catenin signaling represses gene expression, revealing the role of a cis-regulatory motif, the negative regulatory element (NRE). The NRE motif is enriched in the promoters of β-catenin repressed genes and is required for their repression. This provides a comprehensive analysis of the β-catenin-independent arm of the Wnt signaling pathway in a cancer model and suggests that a cis-regulatory grammar may determine Wnt-dependent gene activation versus repression.