Roles of C/EBPα, GATA2, TGF-β-signaling, and epigenetic regulation in the expression of basophil-specific protease genes

Abstract

In the present study, we analyzed the transcriptional regulation of genes encoding basophil-specific proteases Mcpt8 and Mcpt11 to clarify the molecular mechanisms by which the commitment between basophil and mast cell (MC) is determined. We used bone marrow-derived (BM) cells maintained in the presence of IL-3, in which basophil-like cells and MC-like cells exist. Knock down (KD) and overexpression of a transcription factor C/EBP showed that Cebpa mRNA levels were associated with those of Mcpt8 and Prss34 (a gene symbol of Mcpt11). Treatment with trichostatin A (TSA), a histone deacetylase inhibitor, decreased mRNA levels of Cebpa, Mcpt8, and Prss34 in BM cells, whereas these mRNA levels were increased by the treatment with an inhibitor of DNA methyltransferase. TSA treatment upregulated mRNA levels of Gata1 and Mitf in BM cells, and Mitf KD but not Gata1 KD increased mRNA levels of Cebpa and Prss34. Furthermore, Gata2 KD significantly decreased mRNA levels of Mcpt8 and Prss34 without affecting Cebpa mRNA level. In addition, mRNA levels of Gata1 and Mitf were increased in BM cells transfected with Cebpa siRNA, and decreased in Cebpa-overexpressing BM cells. We also found the TGF-{beta} treatment increased mRNA levels of Mcpt8, Prss34, along with upregulation of Gata2 and Cebpa transcription. Taken together, we conclude that the transcription of basophil-specific protease genes was positively regulated by C/EBP, GATA2, and TGF-{beta} signaling with modification of epigenetic regulation.