Article

Genome-wide analysis reveals conserved and divergent features of Notch1/RBPJ binding in human and murine T-lymphoblastic leukemia cells

Department of Pathology, Brigham and Women's Hospital, Boston, MA 02115, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.81). 07/2011; 108(36):14908-13. DOI: 10.1073/pnas.1109023108
Source: PubMed

ABSTRACT Notch1 regulates gene expression by associating with the DNA-binding factor RBPJ and is oncogenic in murine and human T-cell progenitors. Using ChIP-Seq, we find that in human and murine T-lymphoblastic leukemia (TLL) genomes Notch1 binds preferentially to promoters, to RBPJ binding sites, and near imputed ZNF143, ETS, and RUNX sites. ChIP-Seq confirmed that ZNF143 binds to ∼40% of Notch1 sites. Notch1/ZNF143 sites are characterized by high Notch1 and ZNF143 signals, frequent cobinding of RBPJ (generally through sites embedded within ZNF143 motifs), strong promoter bias, and relatively low mean levels of activating chromatin marks. RBPJ and ZNF143 binding to DNA is mutually exclusive in vitro, suggesting RBPJ/Notch1 and ZNF143 complexes exchange on these sites in cells. K-means clustering of Notch1 binding sites and associated motifs identified conserved Notch1-RUNX, Notch1-ETS, Notch1-RBPJ, Notch1-ZNF143, and Notch1-ZNF143-ETS clusters with different genomic distributions and levels of chromatin marks. Although Notch1 binds mainly to gene promoters, ∼75% of direct target genes lack promoter binding and are presumably regulated by enhancers, which were identified near MYC, DTX1, IGF1R, IL7R, and the GIMAP cluster. Human and murine TLL genomes also have many sites that bind only RBPJ. Murine RBPJ-only sites are highly enriched for imputed REST (a DNA-binding transcriptional repressor) sites, whereas human RPBJ-only sites lack REST motifs and are more highly enriched for imputed CREB sites. Thus, there is a conserved network of cis-regulatory factors that interacts with Notch1 to regulate gene expression in TLL cells, as well as unique classes of divergent RBPJ-only sites that also likely regulate transcription.

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    • "It has been observed that some sites in the genome are occupied only when Notch signaling is active (''dynamic sites'') (Bray and Bernard, 2010; Castel et al., 2013; Housden et al., 2013; Krejcí et al., 2009; Wang et al., 2014, 2011). These dynamic sites show increased recoverability by RBP ChIP when NICD is present, leading to the proposal that the NICD/RBP complex acts as a pioneer factor to modify chromatin at these sites (Castel et al., 2013). "
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    • "In support of our hypothesis that NOTCH1 controls the lncRNA transcriptional program in T-ALL, we observed that lncRNAs associated with the top 1,000 most enriched NOTCH1-binding sites were significantly downregulated upon administration of g-SI according to gene set enrichment analysis (GSEA) (Figure 2E). Upon closer examination of many of these lncRNA loci, we observed strong NOTCH1/RBPJk-binding sites (Wang et al., 2011a) at both promoters and intragenic enhancer elements (Figure S2B, highlighted yellow), suggesting direct transcriptional control by Notch signaling. Together these data suggest the presence of a Notch-dependent T-ALL lncRNA expression program, members of which, we believe, may carry important biological functions. "
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