title: Cell-type-specific activation and repression of PU.1 by a complex of discrete, functionally specialized cis-regulatory elements.
abstract: The transcription factor PU.1 is critical for multiple hematopoietic lineages, but different leukocyte types require strictly distinct patterns of PU.1 regulation. PU.1 is required early for T-cell lineage development but then must be repressed by a stage-specific mechanism correlated with commitment. Other lineages require steady, low expression or upregulation. Until now, only the promoter plus a distal upstream regulatory element (URE) could be invoked to explain nearly all Sfpi1 (PU.1) activation and repression, including bifunctional effects of Runx1. However, the URE is dispensable for most Sfpi1 downregulation in early T cells, and we show that it retains enhancer activity in immature T-lineage cells even where endogenous Sfpi1 is repressed. We now present evidence for another complex of conserved noncoding elements that mediate discrete, cell-type-specific regulatory features of Sfpi1, including a myeloid cell-specific activating element and a separate, pro-T-cell-specific silencer element. These elements yield opposite, cell-type-specific responses to Runx1. T-cell-specific repression requires Runx1 acting through multiple nonconsensus sites in the silencer core. These newly characterized sites recruit Runx1 binding in early T cells in vivo and define a functionally specific scaffold for dose-dependent, Runx-mediated repression.
authors.affiliation: Division of Biology, California Institute of Technology, Pasadena, CA 91125, USA.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Zarnegar
authors.forename: Jing
authors.initials: J
authors.lastname: Chen
authors.forename: Ellen V
authors.initials: EV
authors.lastname: Rothenberg
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002454:Cell Differentiation; D002460:Cell Line; D017124:Conserved Sequence; D050676:Core Binding Factor Alpha 2 Subunit / Q000378:metabolism; D004247:DNA / Q000235:genetics; D003851:Deoxyribonucleases / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005816:Genetic Complementation Test; D051379:Mice; D008954:Models, Biological; D008969:Molecular Sequence Data; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism*; D050436:Regulatory Elements, Transcriptional; D012689:Sequence Homology, Nucleic Acid; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism*; D000099232:Proto-Oncogene Protein Spi-1
chemical_list: D050676:Core Binding Factor Alpha 2 Subunit; D004247:DNA; D003851:Deoxyribonucleases; D011518:Proto-Oncogene Proteins; D015534:Trans-Activators; D000099232:Proto-Oncogene Protein Spi-1; C493726:Runx1 protein, mouse