Dissecting the action of an evolutionary conserved non-coding region on renin promoter activity

Nucleic Acids Res. 2007;35(15):5120-9. doi: 10.1093/nar/gkm535. Epub 2007 Jul 26.

Abstract

Elucidating the mechanisms of the human transcriptional regulatory network is a major challenge of the post-genomic era. One important aspect is the identification and functional analysis of regulatory elements in non-coding DNA. Genomic sequence comparisons between related species can guide the discovery of cis-regulatory sequences. Using this technique, we identify a conserved region CNSmd of approximately 775 bp in size, approximately 14 kb upstream of the renin gene. Renin plays a pivotal role for mammalian blood pressure regulation and electrolyte balance. To analyse the cis-regulatory role of this region in detail, we perform 132 combinatorial reporter gene assays in an in vitro Calu-6 cell line model. To dissect the role of individual subregions, we fit several mathematical models to the experimental data. We show that a multiplicative switch model fits best the experimental data and that one subregion has a dominant effect on promoter activity. Mapping of the sub-sequences on phylogenetic conservation data reveals that the dominant regulatory region is the one with the highest multi-species conservation score.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5' Flanking Region
  • Base Sequence
  • Cell Line
  • Conserved Sequence
  • Evolution, Molecular
  • Gene Expression Regulation
  • Genes, Reporter
  • Humans
  • Models, Genetic
  • Promoter Regions, Genetic*
  • Regulatory Elements, Transcriptional*
  • Renin / genetics*

Substances

  • Renin