Isolation of proteins associated with the DNA-bound estrogen receptor alpha.
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.
Schultz-Norton
Jennifer R
JR
Department of Molecular and Integrative Physiology, University of Illinois, Urbana, IL, USA.
Ziegler
Yvonne S
YS
Likhite
Varsha S
VS
Nardulli
Ann M
AM
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
D017931:DNA Primers; D047628:Estrogen Receptor alpha; D011506:Proteins; D004247:DNA