Divergent Binding and Transactivation by Two Related Steroid Receptors at the Same Response Element

J Biol Chem. 2016 May 27;291(22):11899-910. doi: 10.1074/jbc.M115.684480. Epub 2016 Apr 7.

Abstract

Transcription factor (TF) recruitment to chromatin is central to activation of transcription. TF-chromatin interactions are highly dynamic, which are evaluated by recovery half time (t1/2) in seconds, determined by fluorescence recovery experiments in living cells, and chromatin immunoprecipitation (ChIP) analysis, measured in minutes. These two states are related: the larger the t1/2, the longer the ChIP occupancy resulting in increased transcription. Here we present data showing that this relationship does not always hold. We found that histone deacetylase inhibitors (HDACis) significantly increased t1/2 of green fluorescent protein (GFP) fused androgen receptor (AR) on a tandem array of positive hormone response elements (HREs) in chromatin. This resulted in increased ChIP signal of GFP-AR. Unexpectedly, however, transcription was inhibited. In contrast, the GFP-fused glucocorticoid receptor (GR), acting through the same HREs, displayed a profile consistent with current models. We provide evidence that these differences are mediated, at least in part, by HDACs. Our results provide insight into TF action in living cells and show that very closely related TFs may trigger significantly divergent outcomes at the same REs.

Keywords: androgen receptor; chromatin; glucocorticoid receptor; histone deacetylase (HDAC); live-cell dynamics; transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / genetics
  • Adenocarcinoma / metabolism*
  • Aged
  • Animals
  • Chromatin Immunoprecipitation
  • Female
  • Fluorescent Antibody Technique
  • Gene Expression Regulation, Neoplastic*
  • Histone Deacetylase Inhibitors / pharmacology
  • Histone Deacetylases / chemistry
  • Humans
  • Mammary Neoplasms, Animal / genetics
  • Mammary Neoplasms, Animal / metabolism*
  • Mice
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Receptors, Androgen / genetics
  • Receptors, Androgen / metabolism*
  • Receptors, Glucocorticoid / genetics
  • Receptors, Glucocorticoid / metabolism*
  • Response Elements / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcriptional Activation / genetics*
  • Tumor Cells, Cultured

Substances

  • AR protein, mouse
  • Histone Deacetylase Inhibitors
  • RNA, Messenger
  • Receptors, Androgen
  • Receptors, Glucocorticoid
  • Histone Deacetylases

Grants and funding