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Blood, 15 November 2004, Vol. 104, No. 10, pp. 3136-3147.
Prepublished online as a Blood First Edition Paper on August 5, 2004; DOI 10.1182/blood-2004-04-1603.


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Submitted April 30, 2004
Accepted June 22, 2004

Global regulation of erythroid gene expression by transcription factor GATA-1

John J Welch, Jason A Watts, Christopher R Vakoc, Yu Yao, Hao Wang, Ross C Hardison, Gerd A Blobel, Lewis A Chodosh, and Mitchell J Weiss*

Division of Hematology, The Children's Hospital of Philadelphia and the University of Pennsylvania, Philadelphia, PA, USA
Cell and Molecular Biology and Combined Degree Graduate Programs, The University of Pennsylvania School of Medicine, Philadelphia, PA, USA
Department of Biochemistry and Molecular Biology, Center for Comparative Genomics and Bioinformatics, Huck Institutes of Life Sciences,The Pennsylvania State University, University Park, PA, USA
Department of Cancer Biology, Abramson Family Cancer Research Institute, University of Pennsylvania, Philadelphia, PA, USA

* Corresponding author; email: weissmi{at}email.chop.edu.

Transcription factor GATA-1 is required for erythropoiesis, yet its full actions are unknown. We performed transcriptome analysis of G1E-ER4 cells, a GATA-1-null erythroblast line that undergoes synchronous erythroid maturation when GATA-1 activity is restored. We interrogated over 9,000 transcripts at six timepoints representing the transition from late BFU-E to basophilic erythroblast stages. Our findings illuminate several new aspects of GATA-1 function. First, the large number of genes responding quickly to restoration of GATA-1 extends the repertoire of its potential targets. Second, many transcripts were rapidly downregulated, highlighting the importance of GATA-1 in gene repression. Third, upregulation of some known GATA-1 targets was delayed, suggesting that auxiliary factors are required. For example, induction of the direct GATA-1 target gene {beta} major globin was late, and surprisingly, required new protein synthesis. In contrast, the gene encoding Fog1, which cooperates with GATA-1 in {beta} globin transcription, was rapidly induced independently of protein synthesis. Guided by bioinformatic analysis, we demonstrated that selected regions of the Fog1 gene exhibit enhancer activity and in vivo occupancy by GATA-1. These findings define a regulatory loop for {beta} globin expression, and more generally, demonstrate how transcriptome analysis can be used to generate testable hypotheses regarding genetic networks.


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