Direct binding of ZEB2 (Zinc finger E-box-binding homeobo...

Direct binding of ZEB2 (Zinc finger E-box-binding homeobox 2, also known as SIP1) to the E-box elements in the CDH1 gene promoter has been reported in multiple studies. Binding of recombinant mouse Zeb2 to human CDH1 gene promoter was demonstrated by electromobility shift assay (EMSA) in human breast cancer cell line (Comijn et al. 2001) and by chromatin immunoprecipitation (ChIP) in human embryonic kidney cell line HEK293T (van Grunsven et al. 2003) and human epidermoid carcinoma cell line A431 (Mejlvang et al. 2007). The endogenous mouse Zeb2 was shown to bind to the promoter of the endogenous mouse Cdh1 gene in mouse medial-edge epithelium cells in mouse palate by ChIP (Jalali et al. 2012) and a recombinant mouse Zeb2 was shown to bind to the endogenous mouse Cdh1 gene promoter in extracts of mouse mammary NMuMG epithelial cells by EMSA (Shirakihara et al. 2007). Binding of a recombinant human ZEB2 to the endogenous human CDH1 gene promoter in oral squamous carcinoma cell lines was shown by ChIP (Maeda et al. 2005) and binding of a recombinant human ZEB2 to the human CDH1 promoter sequence in vitro was shown by EMSA (Ghoumid et al. 2013). Missense mutations in ZEB2 from Mowat-Wilson syndrome patients abrogate ZEB2 binding to the CDH1 gene promoter (Ghoumid et al. 2013).

ZEB2 binding site in the promoter region of the CDH1 gene partially overlaps with the binding site of KLF4, a transcriptional activator of CDH1. Binding of ZEB2 interferes with KLF4 recruitment, which likely contributes to ZEB2-mediate repression of CDH1 gene transcription (Koopmansch et al. 2013).