In vivo Footprinting analysis of the MnSOD Promoter

With information on chromatin structure we have analyzed hypersensitive site 1 which is superimposed on the MnsOD promoter using in vivo footprinting methodology. This approach provides information on the location of DNA/protein interactions at single nucleotide resolution. The strategy is summarized in the two figures directly below, including the use of ligation-mediated PCR which amplifies the in vivo signal. The figure on the far right below shows an example of the top strand data from position –121 to –254 of the MnSOD promoter.

In vivo footprinting methodology using ligation-mediated PCR (LMPCR) has allowed the detection of 10 basal cis-acting elements in the MnSOD promoter as well as a stimulus-specific protein binding site. The figure below summarizes this data and illustrates the architecture of this TATA- and CAT-less promoter. It also depicts the changes in chromatin structure which ensue following stimulus-specific gene activation.