ERE Project Description:
In chromatin immunoprecipitation experiments, many transcriptional coregulators have been individually shown to be co-localized with estrogen receptor alpha (ERa) on different genomic estrogen response elements (EREs) in an estradiol(E2)-dependent fashion. However, a systematic proteomic analysis of coregulator complexes bound with ERa on EREs has been missing.
Here, we used a biochemical approach - biotinylated ERE-containing DNAs immobilized on streptavidin beads - to pulldown proteins recruited by E2-liganded ERa from MCF-7 and HeLaS3 human cell nuclear extracts, followed by mass spectrometry (MS) or immunoblotting. These experiments utilized two different ERE-containing DNAs, either bearing four (4) copies of the XenopusVitellogenin A2 promoter canonical ERE or a fragment from the human GREB1 gene promoter containing ERE1. Furthermore, nuclear proteins recruited to 4xEREs in the presence of different nucleosomes bearing select histone H3 post-translational modifications, such as H3K4me3 or H3K9me3, were identified.
Standalone ERE Resource Solutions:
Below are FileMaker runtime solutions for Mac OS and Windows that contain the ERE Resource. FileMaker license is not required to use these. The zipped files contain folders with the applications and supporting databases. Please, do NOT delete supporting information from the folders. For more details, refer to the ReadMe file (also available as a page within the ERE Resource Solution).
ReadMe File:
FMP RunTime Solutions:
ERE Resource MS Data Files:
Below are the original mass spectrometry files (Thermo-Fisher instruments, ".raw" format) for the 24 experiments used in construction of the ERE Resource:
