title: Genome-wide analysis reveals conserved and divergent features of Notch1/RBPJ binding in human and murine T-lymphoblastic leukemia cells.
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.
authors: Department of Pathology, Brigham and Women's Hospital, Boston, MA 02115, USA.
authors: Hongfang
authors: H
authors: Wang
authors: James
authors: J
authors: Zou
authors: Bo
authors: B
authors: Zhao
authors: Eric
authors: E
authors: Johannsen
authors: Todd
authors: T
authors: Ashworth
authors: Hoifung
authors: H
authors: Wong
authors: Warren S
authors: WS
authors: Pear
authors: Jonathan
authors: J
authors: Schug
authors: Stephen C
authors: SC
authors: Blacklow
authors: Kelly L
authors: KL
authors: Arnett
authors: Bradley E
authors: BE
authors: Bernstein
authors: Elliott
authors: E
authors: Kieff
authors: Jon C
authors: JC
authors: Aster
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002843:Chromatin / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D015973:Gene Expression Regulation, Leukemic; D015894:Genome, Human; D055106:Genome-Wide Association Study; D006801:Humans; D051819:Immunoglobulin J Recombination Signal Sequence-Binding Protein / Q000235:genetics / Q000378:metabolism*; D015458:Leukemia, T-Cell / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D051881:Receptor, Notch1 / Q000235:genetics / Q000378:metabolism*; D020218:Response Elements; D014158:Transcription, Genetic
chemical_list: D002843:Chromatin; D051819:Immunoglobulin J Recombination Signal Sequence-Binding Protein; C497761:NOTCH1 protein, human; D009363:Neoplasm Proteins; C497762:Notch1 protein, mouse; C497570:RBPJ protein, human; C497571:Rbpj protein, mouse; D051881:Receptor, Notch1