title: Differential binding of the NFE3 and CP1/NFY transcription factors to the human gamma- and epsilon-globin CCAAT boxes.
abstract: Naturally occurring nondeletional mutations affecting the distal CCAAT box of the human gamma-globin gene promoter result in hereditary persistence of fetal hemoglobin in adult life. Although the distal CCAAT box is the target of several factors, including CP1/NFY, CDP, GATA-1 and NFE3, only NFE3 binding activity is consistently sensitive to well characterized mutations in this region such as G-117-->A, C-114-->T, and delta 13 hereditary persistence of fetal hemoglobin. We extensively characterized the binding specificities of NFE3 and demonstrated that NFE3 has unique properties with respect to other CCAAT box-binding proteins. Affinity-purified NFE3 from erythroid K562 cells binds the distal but not the proximal human gamma-globin CCAAT box, the single CCAAT box of the human epsilon-globin promoter, and the proximal CCAAT box of the evolutionarily related Galago crassicaudatus gamma-globin gene. Within the epsilon-globin CCAAT box, NFE3 represents the major and almost exclusive binding activity. Disruption of such a binding site essentially inactivates the epsilon-globin promoter, suggesting that NFE3 plays an important role in the embryonic expression of this gene.
authors.affiliation: Dipartimento di Genetica e di Biologia dei Microrganismi, Università degli Studi di Milano, Italy.
authors.forename: A E
authors.initials: AE
authors.lastname: Ronchi
authors.forename: S
authors.initials: S
authors.lastname: Bottardi
authors.forename: C
authors.initials: C
authors.lastname: Mazzucchelli
authors.forename: S
authors.initials: S
authors.lastname: Ottolenghi
authors.forename: C
authors.initials: C
authors.lastname: Santoro
mesh_terms: D000328:Adult; D001483:Base Sequence; D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D001665:Binding Sites; D022762:CCAAT-Enhancer-Binding Proteins; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005319:Fetal Hemoglobin / Q000235:genetics; D005656:Fungal Proteins / Q000378:metabolism*; D005914:Globins / Q000235:genetics*; D018257:Helix-Loop-Helix Motifs; D006801:Humans; D008745:Methylation; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009687:Nuclear Proteins / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000378:metabolism*; D051837:COUP Transcription Factor II
chemical_list: D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D022762:CCAAT-Enhancer-Binding Proteins; D004268:DNA-Binding Proteins; D005319:Fetal Hemoglobin; D005656:Fungal Proteins; D005914:Globins; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C064042:CBF1 protein, S cerevisiae; C095569:NR2F2 protein, human; D051837:COUP Transcription Factor II