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

doi:10.1093/nar/gkad803

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)

Chemical Biology

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Abstract

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 language	English
Pages (from-to)	10992-11009
Number of pages	18
Journal	Nucleic Acids Research
Volume	51
Issue number	20
DOIs	https://doi.org/10.1093/nar/gkad803
Publication status	Published - 10 Nov 2023

Bibliographical note

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

Funding

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.

Funders	Funder number
Nederlandse Organisatie voor Wetenschappelijk Onderzoek	737.016.014

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Yavuz, S., Kabbech, H., van Staalduinen, J., Linder, S., van Cappellen, W. A., Nigg, A. L., Abraham, T. E., Slotman, J. A., Quevedo, M., Poot, R. A., Zwart, W., van Royen, M. E., Grosveld, F. G., Smal, I., & Houtsmuller, A. B. (2023). Compartmentalization of androgen receptors at endogenous genes in living cells. Nucleic Acids Research, 51(20), 10992-11009. https://doi.org/10.1093/nar/gkad803

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title = "Compartmentalization of androgen receptors at endogenous genes in living cells",

abstract = "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.",

author = "Sel{\c c}uk Yavuz and H{\'e}l{\`e}ne Kabbech and {van Staalduinen}, Jente and Simon Linder and {van Cappellen}, {Wiggert A.} and Nigg, {Alex L.} and Abraham, {Tsion E.} and Slotman, {Johan A.} and Marti Quevedo and Poot, {Raymond A.} and Wilbert Zwart and {van Royen}, {Martin E.} and Grosveld, {Frank G.} and Ihor Smal and Houtsmuller, {Adriaan B.}",

note = "Publisher Copyright: {\textcopyright} The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.",

year = "2023",

month = nov,

day = "10",

doi = "10.1093/nar/gkad803",

language = "English",

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Yavuz, S, Kabbech, H, van Staalduinen, J, Linder, S, van Cappellen, WA, Nigg, AL, Abraham, TE, Slotman, JA, Quevedo, M, Poot, RA, Zwart, W, van Royen, ME, Grosveld, FG, Smal, I & Houtsmuller, AB 2023, 'Compartmentalization of androgen receptors at endogenous genes in living cells', Nucleic Acids Research, vol. 51, no. 20, pp. 10992-11009. https://doi.org/10.1093/nar/gkad803

TY - JOUR

T1 - Compartmentalization of androgen receptors at endogenous genes in living cells

AU - Yavuz, Selçuk

AU - Kabbech, Hélène

AU - van Staalduinen, Jente

AU - Linder, Simon

AU - van Cappellen, Wiggert A.

AU - Nigg, Alex L.

AU - Abraham, Tsion E.

AU - Slotman, Johan A.

AU - Quevedo, Marti

AU - Poot, Raymond A.

AU - Zwart, Wilbert

AU - van Royen, Martin E.

AU - Grosveld, Frank G.

AU - Smal, Ihor

AU - Houtsmuller, Adriaan B.

PY - 2023/11/10

Y1 - 2023/11/10

N2 - 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.

AB - 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.

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U2 - 10.1093/nar/gkad803

DO - 10.1093/nar/gkad803

M3 - Article

C2 - 37791849

AN - SCOPUS:85176509281

SN - 0305-1048

VL - 51

SP - 10992

EP - 11009

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 20

ER -

Compartmentalization of androgen receptors at endogenous genes in living cells

Abstract

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