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Blood, 1 July 2006, Vol. 108, No. 1, pp. 262-269.
Prepublished online as a Blood First Edition Paper on March 7, 2006; DOI 10.1182/blood-2005-07-2878.


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IMMUNOBIOLOGY

FOG-1 represses GATA-1-dependent Fc{epsilon}RI beta-chain transcription: transcriptional mechanism of mast-cell-specific gene expression in mice

Keiko Maeda, Chiharu Nishiyama, Tomoko Tokura, Hiroyasu Nakano, Shunsuke Kanada, Makoto Nishiyama, Ko Okumura, and Hideoki Ogawa

From the Atopy (Allergy) Research Center and Department of Immunology, Juntendo University School of Medicine, Tokyo; and Biotechnology Research Center, The University of Tokyo, Japan.

Cell-type-specific transcription of mouse high-affinity IgE receptor (Fc{epsilon}RI) beta-chain is positively regulated by the transcription factor GATA-1. Although GATA-1 is expressed in erythroid cells, megakaryocytes, and mast cells, the expression of mouse Fc{epsilon}RI beta-chain is restricted to mast cells. In the present study, we characterized the role of GATA-associated cofactor FOG-1 in the regulation of the Fc{epsilon}RI beta-chain promoter. The expression levels of FOG-1, GATA-1, and beta-chain in each hematopoietic cell line were analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting. FOG-1 expression was higher in the beta-chain-negative hematopoietic progenitor cell line Ba/F3 than in the beta-chain-positive mast cell line PT18. By contrast, GATA-1 expression was similar when comparing the 2 cell lines. A transient reporter assay demonstrated that the beta-chain promoter functioned in PT18 but not in Ba/F3 and that the transcription activity of the beta-chain promoter in PT18 was markedly suppressed by overexpression of FOG-1. Although the activity of the beta-chain promoter, which was upregulated by coexpression of GATA-1, was significantly suppressed by coexpression of FOG-1 in the simian kidney CV-1 cells (beta-chain-, GATA-1-, and FOG-1-), the transactivation of the beta-chain promoter by the GATA-1 mutant V205G, which cannot bind FOG-1, was not affected by coexpression of FOG-1. Further, overexpression of FOG-1 in PT18 resulted in decreases in cell surface expression of Fc{epsilon}RI and beta-chain transcription. Finally, suppression of FOG-1 expression using an siRNA approach resulted in increased beta-chain promoter activity in Ba/F3. These results suggest that FOG-1 expression level regulates the GATA-1-dependent Fc{epsilon}RI beta-chain promoter. (Blood. 2006;108:262-269)


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