Huang Duen-Yi, Kuo Yuan-Yeh, Lai Jiann-Shiun, Suzuki Yutaka, Sugano Sumio, Chang Zee-Fen
Graduate Institute of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University, No. 1 Jen Ai Road 1st Section, Taipei, Taiwan, Republic of China.
Nucleic Acids Res. 2004 Jul 27;32(13):3935-46. doi: 10.1093/nar/gkh719. Print 2004.
Expression of Gfi (growth factor-independence)-1B, a Gfi-1-related transcriptional repressor, is restricted to erythroid lineage cells and is essential for erythropoiesis. We have determined the transcription start site of the human Gfi-1B gene and located its first non-coding exon approximately 7.82 kb upstream of the first coding exon. The genomic sequence preceding this first non-coding exon has been identified to be its erythroid-specific promoter region in K562 cells. Using gel-shift and chromatin immunoprecipitation (ChIP) assays, we have demonstrated that NF-Y and GATA-1 directly participate in transcriptional activation of the Gfi-1B gene in K562 cells. Ectopic expression of GATA-1 markedly stimulates the activity of the Gfi-1B promoter in a non-erythroid cell line U937. Interestingly, our results have indicated that this GATA-1-mediated trans-activation is dependent on NF-Y binding to the CCAAT site. Here we conclude that functional cooperation between GATA-1 and NF-Y contributes to erythroid-specific transcriptional activation of Gfi-1B promoter.
Gfi(生长因子独立性)-1B是一种与Gfi-1相关的转录抑制因子,其表达仅限于红系谱系细胞,对红细胞生成至关重要。我们已经确定了人类Gfi-1B基因的转录起始位点,并将其第一个非编码外显子定位在第一个编码外显子上游约7.82 kb处。已确定该第一个非编码外显子之前的基因组序列是其在K562细胞中的红系特异性启动子区域。通过凝胶迁移和染色质免疫沉淀(ChIP)分析,我们证明了NF-Y和GATA-1直接参与K562细胞中Gfi-1B基因的转录激活。GATA-1的异位表达在非红系细胞系U937中显著刺激Gfi-1B启动子的活性。有趣的是,我们的结果表明,这种GATA-1介导的反式激活依赖于NF-Y与CCAAT位点的结合。我们在此得出结论,GATA-1和NF-Y之间的功能合作有助于Gfi-1B启动子的红系特异性转录激活。