By testing promoter activity of ten plant-specific promoters in five eubacterial species we could show that in 50%% of the tested combinations the promoter sequences could be used in the bacterial host [ 12, 13].
This process created proxy datasets for testing promoter occupancy, allowing us to look for documented binding of the TFs in sites overlapping the 4,102 promoters, in the relevant cellular models.
The extent of the effect of the DN-NF-YA, both on NF-Y and chromatin marks binding, is variable on the different tested promoters.
To test promoters, we chose the lens crystallin (Emelyanov and Parinov, 2008) and the sonic hedgehog (shh) promoter (Neumann and Nuesslein-Volhard, 2000).
On all tested promoters the addition of TFEα/β stimulated transcription by approximately twofold to fourfold, with the strong T6 and 16S/23S rRNA promoters showing a weaker response.
Thus, to elucidate which specific genes contribute to the upregulation of miR-16 and miR-26a by p53, we tested promoters of the respective genes in the luciferase reporter assay.
To test promoter activity in rice, the OsNCED3 promoter, encompassing 2407 bp upstream of the ATG start codon of OsNCED3, and the OsNCED4 promoter, a total of 1939 bp upstream of the ATG start codon of OsNCED4, were amplified by polymerase chain reaction (PCR) and fused to a β-glucuronidase (GUS) coding region in the pCAMBIA1305.1 binary vector.
None of the tested promoter constructs showed an increased reporter activity due to c-MYC.
When compared to Nter-15Q, Nter-142Q led to an increase of luciferase activity for all tested promoter regions, with the exception of region A1.
Surprisingly, the expression of the c-MYC protein led to an inhibition of all tested promoter constructs independent of the selected serum concentrations.
