title: Isolation of proteins associated with the DNA-bound estrogen receptor alpha.
abstract: Regulating gene expression is a complex process requiring the interaction of multiple transcription factors with their cognate recognition sequences. While these DNA-bound transcription factors are the primary drivers of gene expression, the capacity of a transcription factor to alter gene expression is tempered by its association with a host of coregulatory proteins that are recruited to the DNA-bound transcription factor. We have developed a novel approach to isolate large complexes of proteins associated with the DNA-bound estrogen receptor alpha (ERalpha) using an agarose-based electrophoretic mobility shift assay (EMSA). This method should be readily adapted to a variety of cultured cell lines, DNA sequences, and transcription factors and has the potential to provide valuable information about a wide variety of regulatory proteins involved in influencing gene expression.
authors.affiliation: Department of Molecular and Integrative Physiology, University of Illinois, Urbana, IL, USA.
authors.forename: Jennifer R
authors.initials: JR
authors.lastname: Schultz-Norton
authors.forename: Yvonne S
authors.initials: YS
authors.lastname: Ziegler
authors.forename: Varsha S
authors.initials: VS
authors.lastname: Likhite
authors.forename: Ann M
authors.initials: AM
authors.lastname: Nardulli
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D017931:DNA Primers; D004587:Electrophoresis, Agar Gel; D024202:Electrophoretic Mobility Shift Assay; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D011506:Proteins / Q000302:isolation & purification* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D017931:DNA Primers; D047628:Estrogen Receptor alpha; D011506:Proteins; D004247:DNA