Conserved elements containing NF-E2 and tandem GATA binding sites are required for erythroid-specific chromatin structure reorganization within the human beta-globin locus control region.
Proper expression of the genes of the human beta-globin gene locus requires the associated locus control region (LCR). Structurally, the LCR is defined by the presence of four domains of erythroid-specific chromatin structure. These domains, which have been characterized as DNase I hypersensitive sites (HSs), comprise the active elements of the LCR. The major focus of this research is to define the cis -acting elements which are required for the formation of these domains of unique chromatin structure. Our previous investigations on the formation of LCR HS4 demonstrated that NF-E2 and tandem, inverted GATA binding sites are required for the formation of the native HS. Similarly arranged NF-E2 and tandem GATA sites are present within the core regions of the other human LCR HSs and are evolutionarily conserved. Using site-directed mutagenesis of human HSs 2 and 3 we have tested the hypothesis that these NF-E2 and GATA sites are common requirements for the formation of all LCR HSs. We find that mutation of these elements, and particularly the GATA elements, results in a decrease or complete loss of DNase I hypersensitivity. These data imply the presence of common structural elements within the core of each LCR HS which are required for erythroid-specific chromatin structure reorganization.
Pomerantz
O
O
Department of Medicine and Department of Pharmacology/Toxicology, Dartmouth Medical School, Hanover, NH 03755, USA.
Goodwin
A J
AJ
Joyce
T
T
Lowrey
C H
CH
D001483:Base Sequence; D002843:Chromatin / Q000235:genetics* / Q000648:ultrastructure; D017124:Conserved Sequence; D004268:DNA-Binding Proteins / Q000235:genetics*; D004912:Erythrocytes / Q000378:metabolism / Q000648:ultrastructure; D050981:Erythroid-Specific DNA-Binding Factors; D050989:GATA2 Transcription Factor; D005914:Globins / Q000235:genetics*; D006454:Hemoglobins / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D050983:NF-E2 Transcription Factor; D050984:NF-E2 Transcription Factor, p45 Subunit; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000235:genetics*
D002843:Chromatin; D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D050989:GATA2 Transcription Factor; C494711:GATA2 protein, human; D006454:Hemoglobins; D050983:NF-E2 Transcription Factor; D050984:NF-E2 Transcription Factor, p45 Subunit; C494692:NFE2 protein, human; D014157:Transcription Factors; D005914:Globins