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Nucleic Acids Research 2004 32(13):3935-3946; doi:10.1093/nar/gkh719
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Published online 27 July 2004

Nucleic Acids Research, Vol. 32 No. 13 © Oxford University Press 2004; all rights reserved

GATA-1 and NF-Y cooperate to mediate erythroid-specific transcription of Gfi-1B gene

Duen-Yi Huang, Yuan-Yeh Kuo, Jiann-Shiun Lai1, Yutaka Suzuki2, Sumio Sugano2 and Zee-Fen Chang*

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, 1 Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan, Republic of China and 2 Laboratory of Genome Structure Analysis, Human Genome Center, the Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minatoku, Tokyo 108-8639, Japan

* To whom correspondence should be addressed. Tel: +886 02 2395 8904; Fax: +886 02 2395 8904; Email: zfchang{at}ha.mc.ntu.edu.tw

Received May 3, 2004; Revised and Accepted July 6, 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 ~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.


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