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Techniques for Evaluation of AR Transcriptional Output and Recruitment to DNA.
Zhang, Manqi; Krause, William C; Agoulnik, Irina U.
Afiliación
  • Zhang M; Department of Chemistry and Biochemistry, Florida International University, Miami, FL, USA.
  • Krause WC; Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA, USA.
  • Agoulnik IU; Biomolecular Sciences Institute, FIU, Miami, FL, USA. iagoulni@fiu.edu.
Methods Mol Biol ; 1786: 219-236, 2018.
Article en En | MEDLINE | ID: mdl-29786796
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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Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Transcripción Genética / Receptores Androgénicos / Regulación de la Expresión Génica / Proteínas de Unión al ADN Límite: Humans / Male Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos

Texto completo: 1 Bases de datos: MEDLINE Asunto principal: Transcripción Genética / Receptores Androgénicos / Regulación de la Expresión Génica / Proteínas de Unión al ADN Límite: Humans / Male Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos