Protein-DNA interactions upstream from the human A gamma globin gene.
We have used DNAase I footprinting and the gel mobility shift assay to study proteins which bind to promoter elements located between -140 and -382 upstream of the human A gamma globin gene. Footprints are found with both erythroid and nonerythroid nuclear extracts at three sites: from -294 to -264, -242 to -227, and -189 to -172 from the transcription initiation site. An erythroid-specific footprint is identified from -194 to -189. We demonstrate that two known transcription factors, the ubiquitous octamer-binding protein OTF-1 and the erythroid regulatory factor NFE-1, bind to the -194 to -172 region and that their footprints overlap. Binding of OTF-1 to this region is reduced by a mutation at -175 associated with a form of non-deletion hereditary persistence of fetal hemoglobin. We conclude that OTF-1 may compete with NFE-1 for the -175 binding site, possibly functioning as a repressor of gamma globin transcription.
O'Neill
D
D
Department of Genetics, Columbia University, New York, NY 10032.
Kaysen
J
J
Donovan-Peluso
M
M
Castle
M
M
Bank
A
A
D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; D005914:Globins / Q000235:genetics*; D051863:Host Cell Factor C1; D006801:Humans; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D050811:Octamer Transcription Factor-1; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D050982:GATA1 Transcription Factor; C497692:HCFC1 protein, human; D051863:Host Cell Factor C1; D050811:Octamer Transcription Factor-1; C494165:POU2F1 protein, human; D014157:Transcription Factors; D005914:Globins; D004247:DNA; D003850:Deoxyribonuclease I