Imagawa S, Yamamoto M, Miura Y
Department of Medicine, Jichi Medical School, Tochigi-ken, Japan.
Acta Haematol. 1996;95(3-4):248-56. doi: 10.1159/000203892.
We examined regulation of the human erythropoietin (Epo) gene through the GATA sequence in the Epo promoter, and demonstrated that Hep3B and HepG2 cells express human GATA-2 (hGATA-2) mRNA and protein. Nuclear extracts of QT6 cells transfected with hGATA-1, -2 or -3 transcription factors revealed specific binding to the GATA element in the human Epo gene promoter by gel mobility shift assay. Transient transfection of Hep3B cells with hGATA-1, -2 or -3 demonstrated that each of these transcription factors significantly decreased the level of expression of Epo mRNA as assessed by a competitive polymerase chain reaction. Furthermore, transient transfection of Hep3B cells with hGATA-1, -2 and -3 and and Epo reporter gene construct showed significant inhibition of the Epo promoter. We conclude that the hGATA-1, -2 and -3 transcription factors specifically bind to the GATA element in the human Epo gene promoter and negatively regulate Epo gene expression.
我们通过促红细胞生成素（Epo）启动子中的GATA序列研究了人类促红细胞生成素（Epo）基因的调控，并证明Hep3B和HepG2细胞表达人类GATA-2（hGATA-2）mRNA和蛋白质。通过凝胶迁移率变动分析，用hGATA-1、-2或-3转录因子转染的QT6细胞的核提取物显示与人类Epo基因启动子中的GATA元件有特异性结合。用hGATA-1、-2或-3对Hep3B细胞进行瞬时转染表明，通过竞争性聚合酶链反应评估，这些转录因子中的每一种都显著降低了Epo mRNA的表达水平。此外，用hGATA-1、-2和-3以及Epo报告基因构建体对Hep3B细胞进行瞬时转染显示对Epo启动子有显著抑制作用。我们得出结论，hGATA-1、-2和-3转录因子特异性结合人类Epo基因启动子中的GATA元件，并对Epo基因表达起负调控作用。
