ZFIN ID: ZDB-PUB-990119-23
Positive and Negative Cis-Acting Elements Are Required for Hematopoietic Expression of Zebrafish GATA-1
Meng, A., Tang, H., Yuan, B.Z., Ong, B.A., Long, Q.M., and Lin, S.
Date: 1999
Source: Blood   93(2): 500-508 (Journal)
Registered Authors: Lin, Shuo, Meng, Anming, Yuan, Ben
Keywords: none
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • DNA-Binding Proteins/genetics*
  • Enhancer Elements, Genetic
  • Erythroid-Specific DNA-Binding Factors
  • GATA1 Transcription Factor
  • Gene Deletion
  • Gene Expression*
  • Gene Expression Regulation, Developmental
  • Green Fluorescent Proteins
  • Hematopoiesis
  • Hematopoietic Stem Cells/metabolism*
  • Humans
  • Luminescent Proteins/genetics
  • Mutagenesis, Site-Directed
  • Point Mutation
  • Promoter Regions, Genetic
  • Recombinant Fusion Proteins
  • Regulatory Sequences, Nucleic Acid*
  • Transcription Factors/genetics*
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish Proteins
PubMed: 9885211
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.