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Blood, Vol. 93 No. 2 (January 15), 1999:
pp. 500-508
Positive and Negative Cis-Acting Elements Are Required for
Hematopoietic Expression of Zebrafish GATA-1
Anming Meng,
Hong Tang,
Baozheng Yuan,
Bruce A. Ong,
Qiaoming Long, and
Shuo Lin
From the Institute of Molecular Medicine and Genetics & Department of
Biochemistry and Molecular Biology, Medical College of Georgia,
Augusta, GA.
GATA-1 is a transcription factor required for development of
erythroid cells. The expression of GATA-1 is tightly restricted to the
hematopoietic lineage. Using transgene constructs containing zebrafish
GATA-1 genomic sequences and the green fluorescent protein (GFP)
reporter gene, we previously showed that a 5.6-kb enhancer/promoter fragment is sufficient to direct erythroid-specific expression of the
GFP. In this study, we used enhancer/promoter fragments containing
various deletion and point mutations to further characterize the
cis-acting elements controlling tissue-specific GATA-1 expression. We
report here the identification of distinct cis-acting elements that
cooperate to confer on GATA-1 its hematopoietic expression pattern. A
CACCC box, located 142 bp upstream of the translation start codon, is
critical for the initiation of GATA-1 expression. A distal double GATA
element is required for maintaining and enhancing the hematopoietic
expression of GATA-1. The erythroid-specific activity of the GATA-1
promoter is also enhanced by a 49-bp sequence element located 218 bp
upstream of the CACCC element and a CCAAT box adjacent to the double
GATA motif. Finally, the hematopoietic specificity of the GATA-1
promoter is secured by a negative cis-acting element that inhibits
expression in the notochord.
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