Structural and functional conservation of the VDREs in primate promoters. A) The nucleotide sequence of each primate VDRE is aligned to demonstrate the high degree of conservation of the direct repeats (upper case) and 3-bp spacer (lower case). C. aethiops contained a G-to-A change in the second direct repeat otherwise all the direct repeats were identical. B) To determine if primate VDREs were activated by 1,25(OH)2D3, the amplified Alus for H. sapiens, M. mulatta and C. aethiops were subcloned into the pGL4 luciferase reporter vector. The constructs were co-transfected into U937 cells with phTKRL (Promega) to control for efficiency. The change in expression is represented as fold-change comparing vehicle treated to 1,25(OH)2D3 treated cells. C) Mononuclear cells were isolated from the peripheral blood of two individual M. mullata and treated with either vehicle or 100 nM 1,25(OH)2D3 for 48 h. The expression level of CAMP mRNA was determined by QRT-PCR and normalized to 18S levels. The data are represented as ng of CAMP per ng of 18S.
Gombart et al. BMC Genomics 2009 10:321 doi:10.1186/1471-2164-10-321