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Identification and Characterization of Androgen Response Elements

  • Reinhilde De Bruyn
  • Rita Bollen
  • Frank ClaessensEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 776)

Abstract

The androgen receptor (AR) has a DNA-binding domain that consists of two zinc coordinating modules. While residues of the first module make most of the sequence-specific contacts, the second module functions as a homodimerization interface (1). This explains why the androgen response elements (AREs) are organized as two 5′-AGAACA-3′-like motifs separated by three basepairs (2). AREs can be located near the promoters of androgen-responsive genes, but are also at considerable distances either upstream or downstream, so the initial steps in locating AREs can be challenging. Traditionally, AR-binding sites were identified by DNA cellulose competition assays (3) or by in vitro footprinting (4). However, the advent of the chromatin immunoprecipitation assays made it possible to identify genomic fragments to which the AR binds either directly or indirectly (5). To enable identification of AREs in such genomic fragments, we developed an in silico approach involving a weight matrix based on all known AREs (6). This will point out candidate AREs, which will still need experimental validation involving a direct interaction assay and a transactivation assay. We describe here the methods most fit to describe an ARE: the electrophoretic mobility shift and the transactivation assays.

Key words

Androgen receptor DNA binding domain androgen response element reporter gene assay electrophoretic mobility shift assay 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Reinhilde De Bruyn
    • 1
  • Rita Bollen
    • 2
  • Frank Claessens
    • 3
    Email author
  1. 1.Department of Molecular Cell Biology and TherapeuticsSt. Jude Children’s Research HospitalMemphisUSA
  2. 2.Department of Molecular Cell BiologyK.U. LeuvenLeuvenBelgium
  3. 3.Molecular Endocrinology Laboratory, Department of Molecular Cell BiologyK.U. LeuvenLeuvenBelgium

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