Analysis of the 5-lipoxygenase promoter and characterization of a vitamin D receptor binding site

Biochim Biophys Acta. 2006 Jul;1761(7):686-97. doi: 10.1016/j.bbalip.2006.04.005. Epub 2006 Apr 26.


1,25-Dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) and transforming growth factor beta (TGFbeta) potently induce 5-lipoxygenase (5-LO) in myeloid cells. We analyzed vitamin D receptor (VDR) binding to putative vitamin D response elements within the 5-LO promoter and analyzed its function by reporter gene analysis. Binding of VDR and retinoid X receptor to the promoter region was shown in DNase I footprinting, electrophoretic mobility shift and chromatin immunoprecipitation assays. However, the identified VDR binding region did not mediate induction of reporter gene activity by 1,25(OH)(2)D(3)/TGFbeta, neither in the 5-LO promoter context nor with the thymidine kinase (tk) promoter. Insertion of the rat atrial natriuretic factor VDRE in reporter plasmids containing the 5-LO promoter diminished induction by 1,25(OH)(2)D(3)/TGFbeta as compared with the tk promoter. Similarly, low inductions were obtained when cells were transiently or stably transfected with constructs containing various 5-LO promoter regions. Concerning basal promoter activity, we identified a positive regulatory region (-779 to -229), which includes the VDR binding region, in 5-LO-positive MonoMac6 cells. In summary, the VDR/RXR complex binds to putative VDREs in the 5-LO promoter, but other sequences outside the 5-LO promoter seem to be responsible or additionally required for the prominent induction of 5-LO mRNA expression by 1,25(OH)(2)D(3) and TGFbeta.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arachidonate 5-Lipoxygenase / genetics*
  • Arachidonate 5-Lipoxygenase / metabolism
  • Binding Sites
  • Calcitriol / metabolism
  • Calcitriol / pharmacology
  • Gene Expression Regulation, Enzymologic / drug effects
  • Genes, Reporter
  • HL-60 Cells
  • Humans
  • Promoter Regions, Genetic*
  • Protein Binding
  • Receptors, Calcitriol / metabolism*
  • Retinoid X Receptors / metabolism
  • Sequence Analysis
  • Sequence Deletion
  • Transforming Growth Factor beta / pharmacology
  • U937 Cells


  • Receptors, Calcitriol
  • Retinoid X Receptors
  • Transforming Growth Factor beta
  • Arachidonate 5-Lipoxygenase
  • Calcitriol