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Interaction between the stage selector element (SSE) in the
proximal
Volume 270,
Number 21,
Issue of May 26, pp. 12892-12898, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
-Globin Gene
-globin promoter and hypersensitivity site 2 in the locus
control region partly mediates the competitive silencing of the
-globin promoter in the fetal developmental stage. We have now
demonstrated that a second SSE-like element in the 5`-untranslated
region of the -gene also contributes to this competitive silencing
of the
-gene. Utilizing transient transfection assays in the fetal
erythroid cell line, K562, we have shown that the core enhancer of
hypersensitivity site 2 can preferentially interact with the proximal
-promoter in the absence of the SSE, completely silencing a linked
-promoter. Mutation of a 20-base pair sequence of the -gene
5`-untranslated region (UTR) led to derepression of
-promoter
activity. A marked activation of -promoter activity was also
observed with this mutation, suggesting the presence of a repressor.
Fine mutagenesis dissected these activities to different regions of the
5`-UTR. The stage selector activity was localized to a region centered
on nucleotides +13 to +15. Electromobility shift assays
utilizing this sequence demonstrated binding of a fetal and
erythroid-specific protein. The repressor activity of the 5`-UTR was
localized to tandem GATA-like sites, which appear to bind a complex of
two proteins, one of which is the erythroid transcription factor
GATA-1. These results indicate that the 5`-UTR of the
-gene
contains sequences that may be important for its transcriptional and
developmental regulation.
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