Abstract
Defining the precise promoter DNA sequence motifs where nuclear receptors and other transcription factors bind is an essential prerequisite for understanding how these proteins modulate the expression of their specific target genes. The purpose of this chapter is to provide the reader with a detailed guide with respect to the materials and the key methods required to perform this type of DNA-binding analysis. Irrespective of whether starting with purified DNA-binding proteins or somewhat crude cellular extracts, the tried-and-true procedures described here will enable one to accurately access the capacity of specific proteins to bind to DNA as well as to determine the exact sequences and DNA contact nucleotides involved. For illustrative purposes, we primarily have used the interaction of the androgen receptor with the rat probasin proximal promoter as our model system.
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Acknowledgments
The authors acknowledge the work of Kimberly J. Reid, Lillian H. Yeung, and Pernille Sorenson in the development of these techniques in our lab.
This work was supported by grants from the Terry Fox Foundation and the National Cancer Institute of Canada.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Read, J.T., Cheng, H., Hendy, S.C., Nelson, C.C., Rennie, P.S. (2009). Receptor-DNA Interactions: EMSA and Footprinting. In: McEwan, I.J. (eds) The Nuclear Receptor Superfamily. Methods in Molecular Biology™, vol 505. Humana Press. https://doi.org/10.1007/978-1-60327-575-0_6
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DOI: https://doi.org/10.1007/978-1-60327-575-0_6
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