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

      Nucleic Acids Research
      5' Untranslated Regions, Base Sequence, Binding Sites, CCAAT-Binding Factor, physiology, DNA-Binding Proteins, Erythroid Cells, metabolism, Erythroid-Specific DNA-Binding Factors, GATA1 Transcription Factor, Humans, K562 Cells, Molecular Sequence Data, Promoter Regions, Genetic, Proto-Oncogene Proteins, biosynthesis, genetics, Repressor Proteins, Transcription Factors, Transcriptional Activation, U937 Cells

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          Abstract

          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.

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