Multiple liver-specific factors bind to a 64-bp element and activate apo(a) gene

Biochem Biophys Res Commun. 2002 Mar 22;292(1):243-9. doi: 10.1006/bbrc.2002.6639.

Abstract

The high plasma levels of lipoprotein(a) [Lp(a)] are associated with atherosclerosis. The apo(a) gene is responsible for the variance of Lp(a) concentration and its expression is liver-specific. By 5'-deletion analysis, we, in a luciferase gene reporter assay, have identified a 64-bp AT-rich region of upstream apo(a) gene (-703 to -640) that binds to multiple liver-specific factors. The 64 bp cis-element contained three dyad symmetry elements (DSEs) that are crucial for synergistic binding to the factors. We have demonstrated that both DSE-2 and -3 together are responsible for factor binding in vitro, and for gene activation in liver cells. Further, we have purified one of the UV cross-linked DNA-protein complexes to homogeneity by streptavidin magnetic bead chromatography. The identification of a further upstream negative regulatory region (-1432 to -704) led us to predict that as yet unidentified transcriptional repressor(s) might also repress apo(a) gene transcription.

MeSH terms

  • AT Rich Sequence
  • Apolipoproteins A / genetics*
  • Base Sequence
  • Cell Line
  • DNA-Binding Proteins / isolation & purification
  • DNA-Binding Proteins / metabolism
  • HeLa Cells
  • Humans
  • Liver / metabolism*
  • Luciferases / analysis
  • Molecular Sequence Data
  • Promoter Regions, Genetic
  • Response Elements*
  • Sequence Deletion
  • Simian virus 40 / genetics
  • Streptavidin / chemistry
  • Trans-Activators / isolation & purification
  • Trans-Activators / metabolism*
  • Transcriptional Activation*

Substances

  • Apolipoproteins A
  • DNA-Binding Proteins
  • Trans-Activators
  • Streptavidin
  • Luciferases