The role of the general transcription factor IIF in androgen receptor-dependent transcription

Molecular Endocrinology
M Ansar ChoudhryIain J McEwan

Abstract

The androgen receptor (AR) is a member of the steroid receptor subfamily of nuclear receptors and is important for normal male sexual differentiation and fertility. The major transactivation function of the AR, termed activation function 1 (AF1), is modular in structure and has been mapped to the N terminus of the protein. To understand better the mechanisms whereby the AR activates transcription, we have established a novel cell-free transcription assay. This is based on the use of a dual reporter gene template, containing promoter proximal and distal G-less cassettes, which result in different size transcripts that can be easily detected and quantified. The promoter proximal transcript gives an indication of transcription initiation and promoter escape, whereas the relative levels of the distal transcript indicate elongation efficiency. The AR-AF1-Lex protein enhanced production of both transcripts whereas, in the absence of DNA binding, the AF1 domain squelched both initiation and elongation. Mutations in the transactivation domain that impaired transactivation and/or binding of the general transcription factor IIF (TFIIF) were found to reduce the ability of AR-AF1 to squelch transcription. Addition of recombinant TFIIF reve...Continue Reading

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Citations

Mar 24, 2010·Molecular Endocrinology·Tushar MenonDaniel A Bochar
Aug 2, 2011·Molecular and Cellular Endocrinology·Krista K HillDean P Edwards
Jun 26, 2013·Molecular and Cellular Endocrinology·S Stoney Simons, Raj Kumar
Nov 29, 2013·Molecular Endocrinology·S Stoney SimonsRaj Kumar

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