The K442A R447A mutant lacks two key side chains critical for sequence-specific DNA recognition by GR at multiple response elements (refs. 13,19, and Supplementary Figure 2c).
An additional level of complexity is added by the frequent presence of multiple response elements in gene promoter regions.
NFκB reporter plasmid consisted of multiple response elements upstream of destabilized green fluorescent protein gene that encodes for d2EGFP reporter protein [39].
The backbone vector (p-luc in Fig. 1) produced no significant luciferase signal above autofluorescent background upon treatment with androgen, while activity of a positive control reporter vector (MMTV-luc, containing multiple response elements) was induced eight- to tenfold (data not shown).
The Cignal Lenti C/EBP reporter, which expresses a luciferase gene driven by multiple transcription response element (ATTGCGCAAT) repeats, was obtained from SABiosciences (Frederick, MD, USA).
To evaluate how proteasome inhibitors affect FoxM1 transcriptional activity we used a derivative of U2OS osteosarcoma cell line, C3-Luc cells with a doxycycline-inducible FoxM1-GFP fusion protein and a firefly luciferase under the control of multiple FoxM1 response elements [13].
Estrogen may be an important regulator, as multiple estrogen response elements (EREs) within the human LKB1 promoter region confer a repressive action in estrogen receptor (ER -positive MCF-7 human brER -positivecells [ 29].
We have also shown that DDT activates transcription at multiple DNA response elements through p38-mediated phosphorylation and activation of the coactivators p300 (Bratton et al. 2009) and GRIP1 (Frigo et al. 2006).
The trans-activation potencies of MEHP, Wy-14,643, dichloroacetic acid (DCA), and TCA for the mouse PPAR-α were monitored using a luciferase reporter gene containing multiple PPAR response elements derived from the rat hydratase/dehydrogenase promoter in transiently transfected COS-1 monkey kidney cells.
We identified multiple consensus progesterone response element (PRE) sequences up- and downstream of the MSX2 transcriptional start site using MatInspector software [ 51].
