Positive and negative cis-acting elements are required for hematopoietic expression of zebrafish GATA-1

• Published in: Blood Vol. 93; no. 2; pp. 500 - 508
• Main Authors: ANMING MENG, HONG TANG, BAOZHENG YUAN, ONG, B. A, QIAOMING LONG, SHUO LIN
• Format: Journal Article
• Language: English
• Published: Washington, DC The Americain Society of Hematology 15.01.1999
• Subjects: Animals; Animals, Genetically Modified; Biological and medical sciences; Cell differentiation, maturation, development, hematopoiesis; Cell physiology; DNA-Binding Proteins - genetics; Enhancer Elements, Genetic; Erythroid-Specific DNA-Binding Factors; Fundamental and applied biological sciences. Psychology; GATA1 Transcription Factor; Gene Deletion; Gene Expression; Gene Expression Regulation, Developmental; Green Fluorescent Proteins; Hematopoiesis; Hematopoietic Stem Cells - metabolism; Humans; Luminescent Proteins - genetics; Molecular and cellular biology; Mutagenesis, Site-Directed; Point Mutation; Promoter Regions, Genetic; Recombinant Fusion Proteins; Regulatory Sequences, Nucleic Acid; Transcription Factors - genetics; Zebrafish - embryology; Zebrafish - genetics; Zebrafish Proteins; Vertebrata; Hematopoiesis; Stem cell; Pisces; Amphibia; Transgenic animal; Hematopoietic cell; Salientia; Gene expression; Transcription factor GATA1; Cis stereoisomer
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.v93.2.500

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.