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• W2079073757 abstract "Thrombopoietin (TPO), the ligand for c-Mpl, is a cytokine that regulates megakaryocyte growth and development. We have cloned the 5′-flanking region of the human TPO gene and analyzed its promoter activity. The human TPO gene promoter lacks a TATA box and directs transcription initiation at multiple sites over a 50-nucleotide region. Transient expression in a human liver cell line (PLC) of promoter fragment-luciferase reporter gene constructs containing a series of 5′-truncated sequences or site-directed mutations identified a sequence 5′-ACTTCCG-3′ from −69 to −63 as a positive cis-acting element for high level expression of TPO gene. This sequence contains a core motif (C/A)GGA(A/T) for Ets family proteins in the noncoding strand. Gel mobility shift assays performed with nuclear protein from PLC cells identified a DNA binding protein(s) specific for the element. Anti-E4TF1-60(GABPα) or anti-E4TF1-53/47(GABPβ) antibodies supershifted the complex in gel shift assay. Furthermore, co-expression of E4TF1-60 and E4TF1-53/47 squelched TPO gene expression in PLC and HepG2 cells. It is concluded that Ets family transcription factor E4TF1(GABPα/β), an ubiquitously expressed protein, is required for high level expression of the TPO gene in liver. Thrombopoietin (TPO), the ligand for c-Mpl, is a cytokine that regulates megakaryocyte growth and development. We have cloned the 5′-flanking region of the human TPO gene and analyzed its promoter activity. The human TPO gene promoter lacks a TATA box and directs transcription initiation at multiple sites over a 50-nucleotide region. Transient expression in a human liver cell line (PLC) of promoter fragment-luciferase reporter gene constructs containing a series of 5′-truncated sequences or site-directed mutations identified a sequence 5′-ACTTCCG-3′ from −69 to −63 as a positive cis-acting element for high level expression of TPO gene. This sequence contains a core motif (C/A)GGA(A/T) for Ets family proteins in the noncoding strand. Gel mobility shift assays performed with nuclear protein from PLC cells identified a DNA binding protein(s) specific for the element. Anti-E4TF1-60(GABPα) or anti-E4TF1-53/47(GABPβ) antibodies supershifted the complex in gel shift assay. Furthermore, co-expression of E4TF1-60 and E4TF1-53/47 squelched TPO gene expression in PLC and HepG2 cells. It is concluded that Ets family transcription factor E4TF1(GABPα/β), an ubiquitously expressed protein, is required for high level expression of the TPO gene in liver." @default.
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• W2079073757 date "1997-04-01" @default.
• W2079073757 modified "2023-10-18" @default.
• W2079073757 title "Characterization of the Human Thrombopoietin Gene Promoter" @default.
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• W2079073757 doi "https://doi.org/10.1074/jbc.272.17.11361" @default.
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