Genomic organization, promoter cloning, and chromosomal localization of the Dif-2 gene.
We describe the genomic organization and the functional promoter of the monocyte specific gene Dif-2, the human homologue to genes in mouse (gly96) and rat (PRG1), that is downregulated during cell differentiation. The Dif-2 gene consists of two exons and a single intron of 112 bp in length. RNase protection assay indicates one major transcription start site. Sequence analysis reveals several consensus sequences for transcription factors including NF-kappa B, C/EBP, SP1, and the lack of a classical TATA-box. To demonstrate promoter activity, DNA fragments of the Dif-2 5'-flanking region were ligated upstream to the luciferase gene and transfected into HepG2 and HeLa cells. A minimal promoter element between nt -158 and nt +74 containing NF-kappa B and SP1 binding sites was shown to be sufficient for basal activity. These transcription factor binding sites, which are conserved between Dif-2, gly96, and PRG1 promoter regions, indicate a significant role for Dif-2 expression and may explain LPS and C2-ceramide sensitivity. The Dif-2 gene was mapped to chromosome 6p21.3 using in situ hybridization technique.
Pietzsch
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A
Institute for Clinical Chemistry, University of Regensburg, Germany.
Büchler
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C
Schmitz
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G
D000818:Animals; D051017:Apoptosis Regulatory Proteins; D001483:Base Sequence; D001665:Binding Sites; D002454:Cell Differentiation; D002896:Chromosomes, Human, Pair 6; D003001:Cloning, Molecular; D016384:Consensus Sequence; D015536:Down-Regulation; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D017874:Immediate-Early Proteins; D017404:In Situ Hybridization, Fluorescence; D008156:Luciferases / Q000378:metabolism; D008565:Membrane Proteins; D051379:Mice; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins; D010750:Phosphoproteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051381:Rats; D012260:Ribonucleases / Q000378:metabolism; D014158:Transcription, Genetic
D051017:Apoptosis Regulatory Proteins; D006023:Glycoproteins; C099815:IER3 protein, human; D017874:Immediate-Early Proteins; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D010750:Phosphoproteins; D008156:Luciferases; D012260:Ribonucleases