The Subnuclear Three-dimensional Image Analysis of Androgen Receptor Fused to Green Fluorescence Protein

Arihiro Tomura, Kiminobu Goto, Hidetaka Morinaga, Masatoshi Nomura, Taijiro Okabe, Toshihiko Yanase, Ryoichi Takayanagi, Hajime Nawata

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Abstract

To establish the novel approach in order to distinguish the transcriptionally active androgen receptor (AR) from the transcriptionally inactive AR, we performed the three-dimensional construction of confocal microscopic images of intranuclear AR. This method clearly distinguished the subnuclear localization of transcriptionally active AR tagged with green fluorescent protein (AR-GFP) from the transcriptionally inactive AR-GFP. Transcriptionally active AR-GFP mainly produced 250-400 fluorescence foci in the boundary region between euchromatin and heterochromatin. Although the AR-GFP bound to such antiandrogens as hydroxyflutamide or bicalutamide translocated to the nucleus, they homogeneously spread throughout the nucleus without producing any fluorescence foci. Antiandrogenic environmental disrupting chemicals, such as 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene, vinclozolin, or nitrofen, also disrupted the intranuclear fluorescence foci. A point mutation (T877A) resulted in the loss of ligand specificity in AR-GFP. Even in this mutant receptor, agonists, such as dihydrotestosterone, hydroxyflutamide, or progesterone, produced the fluorescence foci in the nucleus, whereas the transcriptionally inactive mutant binding bicalutamide was observed to be spread homogeneously in the nucleus. Taken together, our findings suggest that, after nuclear translocation, AR is possibly located in the specific region in the nucleus while demonstrating clustering tightly depending on the agonist-induced transactivation competence.

Original languageEnglish
Pages (from-to)28395-28401
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number30
DOIs
Publication statusPublished - Jul 27 2001

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All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Tomura, A., Goto, K., Morinaga, H., Nomura, M., Okabe, T., Yanase, T., ... Nawata, H. (2001). The Subnuclear Three-dimensional Image Analysis of Androgen Receptor Fused to Green Fluorescence Protein. Journal of Biological Chemistry, 276(30), 28395-28401. https://doi.org/10.1074/jbc.M101755200