Dissecting the action of an evolutionary conserved non-coding region on renin promoter activity.
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.
Institute for Physiology, Systems Biology-Computational Physiology, Charité Universitätsmedizin, Berlin. ralf.mrowka@charite.de
Ralf
R
Mrowka
Andreas
A
Steege
Charlotte
C
Kaps
Hanspeter
H
Herzel
Bernd J
BJ
Thiele
Pontus B
PB
Persson
Nils
N
Blüthgen
D024506:5' Flanking Region; D001483:Base Sequence; D002460:Cell Line; D017124:Conserved Sequence; D019143:Evolution, Molecular; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D008957:Models, Genetic; D011401:Promoter Regions, Genetic; D050436:Regulatory Elements, Transcriptional; D012083:Renin / Q000235:genetics*
D012083:Renin