Compartmentalization of androgen receptors at endogenous genes in living cells

Selçuk Yavuz, Hélène Kabbech, Jente van Staalduinen, Simon Linder, Wiggert A. van Cappellen, Alex L. Nigg, Tsion E. Abraham, Johan A. Slotman, Marti Quevedo, Raymond A. Poot, Wilbert Zwart, Martin E. van Royen, Frank G. Grosveld, Ihor Smal, Adriaan B. Houtsmuller (Corresponding author)

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A wide range of nuclear proteins are involved in the spatio-temporal organization of the genome through diverse biological processes such as gene transcription and DNA replication. Upon stimulation by testosterone and translocation to the nucleus, multiple androgen receptors (ARs) accumulate in microscopically discernable foci which are irregularly distributed in the nucleus. Here, we investigated the formation and physical nature of these foci, by combining novel fluorescent labeling techniques to visualize a defined chromatin locus of AR-regulated genes-PTPRN2 or BANP-simultaneously with either AR foci or individual AR molecules. Quantitative colocalization analysis showed evidence of AR foci formation induced by R1881 at both PTPRN2 and BANP loci. Furthermore, single-particle tracking (SPT) revealed three distinct subdiffusive fractional Brownian motion (fBm) states: immobilized ARs were observed near the labeled genes likely as a consequence of DNA-binding, while the intermediate confined state showed a similar spatial behavior but with larger displacements, suggesting compartmentalization by liquid-liquid phase separation (LLPS), while freely mobile ARs were diffusing in the nuclear environment. All together, we show for the first time in living cells the presence of AR-regulated genes in AR foci.

Original languageEnglish
Pages (from-to)10992-11009
Number of pages18
JournalNucleic Acids Research
Issue number20
Publication statusPublished - 10 Nov 2023

Bibliographical note

Publisher Copyright:
© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.


Funding Dutch Research Council (NWO) through the Building Blocks of Life program (GENOMETRACK project) [737.016.014]. Funding for open access charge: Erasmus MC, Rotterdam, The Netherlands.

FundersFunder number
Nederlandse Organisatie voor Wetenschappelijk Onderzoek737.016.014


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