Genomic Organization, Promoter Cloning, and Chromosomal Localization of the Dif-2 Gene

doi:10.1006/bbrc.1998.8500

Biochemical and Biophysical Research Communications

Volume 245, Issue 3, 28 April 1998, Pages 651-657

https://doi.org/10.1006/bbrc.1998.8500 Get rights and content

Abstract

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-κB, 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-κB 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 usingin situhybridization technique.

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^☆: The novel nucleotide sequences published here have been submitted to the EMBL sequence data bank and are available under accession numbers Y16736 and Y16737.

^☆☆: Abbreviations used: C/EBP, CCAAT/enhancer binding protein; FISH, fluorescencein situhybridization; NF-κB, nuclear factor-κB; LPS, lipopolysaccharide; lysoPC, lysophosphatidylcholine; M-CSF, macrophage-colony stimulating factor; PMA, phorbol-12-myristate-13-acetate; USF, upstream stimulatory factor

²: To whom correspondence should be addressed at Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Franz-Josef-Strauß-Allee 11, D-93053 Regensburg, Germany. Fax: +49-941-9446202. E-mail:[email protected].

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Regular ArticleGenomic Organization, Promoter Cloning, and Chromosomal Localization of the Dif-2 Gene☆,☆☆

Abstract

Curr. Opin. Immunol.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Anal. Biochem.

J. Biol. Chem.

Cell

Biochim. Biophys. Acta

Trends Biochem. Sci.

Cell

FEBS Lett.

J. Leukoc. Biol.

Regular Article
Genomic Organization, Promoter Cloning, and Chromosomal Localization of the Dif-2 Gene☆,☆☆