Abstract
The FcR beta-chain, a subunit of two related multisubunit receptor complexes, the FcepsilonRI and FcgammaRIII, amplifies the mast cell response and is necessary for the cell surface expression of FcepsilonRI in mouse. The transient reporter assay indicated that -69/+4 region is required for cell type-specific transcriptional regulation of mouse beta-chain gene. EMSA using Abs against transcription factors or competitive oligonucleotides demonstrated that -58/-40 region (containing overlapping three GATA-1 sites, -53/-48, -46/-51, and -42/-47) and -31/-26 region (containing one GATA-1 site) are recognized by GATA-1. The promoter activity of beta-chain was decreased by nucleotide replacements of the GATA-1 sites in mouse mast cell line PT18. Furthermore, exogenously produced GATA-1 up-regulated the promoter activity in CV-1 cells, which are negative in the beta-chain production and the up-regulation was apparently suppressed by GATA-1 site mutations. These results indicate that cell type-specific transcription of mouse beta-chain gene is regulated by GATA-1.
Highlights
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The luciferase activity in the PT18 cells which were transiently transfected with Ϫ2.4k/pGL3-Basic was ϳ10-fold higher than that from PT18 cells transfected with pGL3-Basic, whereas no difference was observed between the luciferase activities directed by Ϫ2.4k/pGL3-Basic and that of pGL3-Basic when the reporter plasmids were introduced into Fc⑀RI -chain negative cell lines A20.2J, L5178Y, or RAW264.7 (Fig. 1)
We indicated that the mouse -chain promoter is functional only in PT18 mouse mast cell line, but not in monomacrophage and lymphoma cell lines
Summary
Keiko Maeda,* Chiharu Nishiyama,2* Tomoko Tokura,* Yushiro Akizawa,* Makoto Nishiyama,† Hideoki Ogawa,* Ko Okumura,* and Chisei Ra*‡ The transient reporter assay indicated that ؊69/؉4 region is required for cell type-specific transcriptional regulation of mouse -chain gene.
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