Genomic organization, promoter cloning, and chromosomal localization of the Dif-2 gene

Biochem Biophys Res Commun. 1998 Apr 28;245(3):651-7. doi: 10.1006/bbrc.1998.8500.

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-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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis Regulatory Proteins
  • Base Sequence
  • Binding Sites
  • Cell Differentiation
  • Chromosomes, Human, Pair 6*
  • Cloning, Molecular
  • Consensus Sequence
  • Down-Regulation
  • Glycoproteins / genetics*
  • Humans
  • Immediate-Early Proteins*
  • In Situ Hybridization, Fluorescence
  • Luciferases / metabolism
  • Membrane Proteins
  • Mice
  • Molecular Sequence Data
  • Neoplasm Proteins*
  • Phosphoproteins / genetics*
  • Promoter Regions, Genetic*
  • Rats
  • Ribonucleases / metabolism
  • Transcription, Genetic

Substances

  • Apoptosis Regulatory Proteins
  • Glycoproteins
  • IER3 protein, human
  • Immediate-Early Proteins
  • Membrane Proteins
  • Neoplasm Proteins
  • Phosphoproteins
  • Luciferases
  • Ribonucleases

Associated data

  • GENBANK/Y16736
  • GENBANK/Y16737