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Receptor-DNA interactions: EMSA and footprinting

By Jason T. Read, Helen Cheng, Stephen C. Hendy, Colleen C. Nelson and Paul S. Rennie

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

Topics: 060100 BIOCHEMISTRY AND CELL BIOLOGY, 060407 Genome Structure and Regulation, 110105 Medical Biochemistry - Nucleic Acids, 111201 Cancer Cell Biology, Nuclear Extracts, Methylation Interference, Androgen Receptor, Probasin Promoter, EMSA
Publisher: Humana
Year: 2009
DOI identifier: 10.1007/978-1-60327-575-0_6
OAI identifier: oai:eprints.qut.edu.au:37405
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