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Mol. Cell. Biol., May 1995, 2437-2447, Vol 15, No. 5
Copyright © 1995, American Society for Microbiology

Functional synergy and physical interactions of the erythroid transcription factor GATA-1 with the Kruppel family proteins Sp1 and EKLF

M Merika and SH Orkin
Division of Hematology-Oncology, Children's Hospital, Boston, Massachusetts 02115, USA.

An unresolved aspect of current understanding of erythroid cell- specific gene expression relates to how a limited number of transcriptional factors cooperate to direct high-level expression mediated by cis-regulatory elements separated over large distances within globin loci. In this report, we provide evidence that GATA-1, the major erythroid transcription factor, activates transcription in a synergistic fashion with two Kruppel family factors, the ubiquitous protein Sp1 and the erythroid-restricted factor EKLF (erythroid Kruppel- like factor), which recognize GC and/or GT/CACC motifs. Binding sites for both GATA-1 and these Kruppel proteins (especially Sp1) are found in close association in the promoters and enhancers of numerous erythroid cell-expressed genes and appear to cooperate in directing their expression. We have shown that GATA-1 interacts physically with Sp1 and EKLF and that interactions are mediated through their respective DNA-binding domains. Moreover, we show that GATA-1 and Sp1 synergize from a distance in constructs designed to mimic the architecture of globin locus control regions and downstream globin promoters. Finally, the formation of GATA-1-SP1 complexes was demonstrated in vivo by the ability of Sp1 to recruit GATA-1 to a promoter in the absence of GATA-binding sites. These experiments provide the first evidence for functionally important protein-protein interactions involved in erythroid cell-specific expression and suggest a mechanism by which DNA loops between locus control regions and globin promoters (or enhancers) might be formed or stabilized.


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