Analysis of possible repressor elements in the 5'-flanking region of the human beta-globin gene.
Human beta-globin gene expression is confined predominantly to the adult with little or no expression of this gene occurring during embryonic or fetal life. The lack of expression of this gene in embryonic and fetal erythroid tissue could be due to the absence of required positive regulatory factors in these cells or the presence of negative regulatory factors which prevent expression of the adult globin gene. To test the repressor model, we have used a gel electrophoretic mobility shift assay to identify regions in the human beta-globin gene which bind proteins found in K562 cells, a cell line that expresses embryonic and fetal globins but not adult beta-globin. DNA fragments comprising the entire human beta-globin gene were assayed using nuclear proteins from K562 cells, and four regions were found that bind proteins. These are located within the 5'-flanking region, within the first and second introns, and at the 3'-flanking region of the gene. Previous studies have suggested the presence of potential repressor sites 5' of exon 2. For this reason, we examined whether the lack of the binding regions in the 5'-flanking sequence allow expression of the human beta-globin gene in transgenic mice during embryonic life. beta-globin gene expression was confined to adult life, indicating that if a transcriptional repressor is responsible for inactivating this gene in embryonic tissue, it is not regulated solely by sequences upstream from -122 bp in the 5'-flanking region of the human beta-globin gene.
Krakowsky
J M
JM
Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati, College of Medicine 45267-0524.
Panke
E S
ES
Lee
R F
RF
McNeish
J
J
Potter
S S
SS
Lingrel
J B
JB
D000818:Animals; D004252:DNA Mutational Analysis; D005786:Gene Expression Regulation; D005809:Genes, Regulator / Q000502:physiology*; D005914:Globins / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D012045:Regulatory Sequences, Nucleic Acid; D015183:Restriction Mapping
D005914:Globins