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Techniques for Evaluation of AR Transcriptional Output and Recruitment to DNA

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Prostate Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1786))

Abstract

Steroid receptors are ligand activated transcription factors whose promoter specificity is regulated by a broad set of coregulators and pioneer factors. Corepressors and coactivators determine receptors’ recruitment to specific regulatory elements and ultimately their transcriptional output. Using androgen receptor (AR) and NCOR1 corepressor as examples, this chapter describes experimental approaches to evaluate recruitment of steroid receptors and their coregulators to DNA and to determine coregulator contribution to the transcriptional output of the receptor. The chromatin immunoprecipitation assay, or ChIP, quantifies protein–DNA interaction in the cellular chromatin environment. Here, we describe a protocol to measure NCOR1 recruitment to AR binding sites of interest using ChIP. Gene Set Enrichment Analysis, GSEA, is a computational technique to determine whether a defined gene set is significantly represented among changes in gene expression between two biological groups. As an example, we examine whether AR repressed genes are significantly represented among genes altered by the NCOR1 knockout.

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Author information

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Florida International University, Miami, FL, USA

    Manqi Zhang

  2. Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA, USA

    William C. Krause

  3. Biomolecular Sciences Institute, FIU, Miami, FL, USA

    Irina U. Agoulnik

  4. Baylor College of Medicine, Houston, TX, USA

    Irina U. Agoulnik

  5. Department of Human and Molecular Genetics, Herbert Wertheim College of Medicine, Miami, FL, USA

    Irina U. Agoulnik

Authors
  1. Manqi Zhang
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  2. William C. Krause
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  3. Irina U. Agoulnik
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Corresponding author

Correspondence to Irina U. Agoulnik .

Editor information

Editors and Affiliations

  1. Department of Urology, Innsbruch Medical University, Innsbruck, Austria

    Zoran Culig

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Zhang, M., Krause, W.C., Agoulnik, I.U. (2018). Techniques for Evaluation of AR Transcriptional Output and Recruitment to DNA. In: Culig, Z. (eds) Prostate Cancer. Methods in Molecular Biology, vol 1786. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7845-8_13

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  • DOI: https://doi.org/10.1007/978-1-4939-7845-8_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7843-4

  • Online ISBN: 978-1-4939-7845-8

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