The androgen receptor depends on ligand‐binding domain dimerization for transcriptional activation

Kharraz, Sarah El; Dubois, Vanessa; Royen, Martin E. van; Houtsmuller, Adriaan B.; Pavlova, Ekaterina; Atanassova, Nina; Nguyen, Tien T.; Voet, Arnout; Eerlings, Roy; Handle, Florian; Prekovic, Stefan; Smeets, Elien; Moris, Lisa; Devlies, Wout; Ohlsson, Claes; Poutanen, Matti; Verstrepen, Kevin J.; Carmeliet, Geert; Launonen, Kaisa-Mari; Helminen, Laura; Palvimo, Jorma J.; Libert, Claude; Vanderschueren, Dirk; Helsen, Christine; Claessens, Frank

doi:10.15252/embr.202152764

Cited by 30 publications

(19 citation statements)

References 63 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The downregulation of Rhox5 expression ( Fig 7 ), which confirms our previous findings [8], further suggests that AR signaling may also be impaired in Sertoli cells. Recently, in a mouse model with disrupted dimerization of the AR ligand-binding-domain, a decreased expression of Hsd17b3 and of AR-regulated genes, such as Rhox5 and Insl3 , a higher percentage of Sertoli cells and a decreased production of round and elongated spermatids were found [46], which further supports our hypothesis that AR signaling may be defective in organotypic cultures. In the SPARKI (SPecificity-affecting AR KnockIn)-AR mouse model, which has a defect in binding to AR-specific DNA motifs while displaying normal interaction with the classical androgen response elements (AREs), it has been shown that Rhox5 expression is regulated by an AR-selective ARE, while Eppin expression is regulated by a classical, nonselective ARE [47].…”

Section: Discussionsupporting

confidence: 85%

Steroidogenesis and androgen/estrogen signaling pathways are altered inin vitromatured testicular tissues of prepubertal mice

Moutard

Goudin

Jaeger

et al. 2022

Preprint

View full text Add to dashboard Cite

Cancer treatments such as chemotherapy can have gonadotoxic effects. In order to preserve and restore the fertility of prepubertal patients with cancer, testicular biopsies are frozen and could theoretically be later matured in vitro to produce spermatozoa for assisted reproductive technology. A complete in vitro spermatogenesis has been obtained from prepubertal testicular tissue in the mouse model, although the sperm yield was low. Since steroid hormones play an essential role in spermatogenesis, it appears necessary to ensure that their synthesis and mechanisms of action are not altered in in vitro cultured tissues. The aim of this study was therefore to investigate steroidogenesis as well as androgen and estrogen signaling during in vitro maturation of mouse prepubertal testicular tissues. Histological, RT-qPCR, Western blot analyses, measurements of cholesterol, steroid hormones levels and aromatase activity were performed on fresh or frozen/thawed in vitro cultured mouse testicular tissues from 6.5 days postpartum (dpp) mice as well as on age-matched in vivo controls. A similar density of Leydig cells (LC) was found after 30 days of organotypic culture (D30) and at 36.5 dpp, the corresponding in vivo time point. However, LC were partially mature after in vitro culture, with decreased Sult1e1 and Insl3 mRNA levels (adult LC markers). Moreover, the transcript levels of Cyp11a1, Cyp17a1 and Hsd17b3 encoding steroidogenic enzymes were decreased in vitro. Increased amounts of progesterone and estradiol and reduced androstenedione intratesticular levels were observed at D30. Furthermore, androgen signaling was altered at D30, with decreased transcript levels of androgen target genes (Rhox5, Septin12). Moreover, the expression and activity of aromatase and estrogen signaling were impaired at D30. The addition of hCG to the organotypic culture medium induced an elevation in androgen production but did not improve sperm yield. In conclusion, this study reports partial LC maturation, disturbed steroidogenic activity of LC, abnormal steroid hormone content as well as altered androgen and estrogen signaling in cultures of fresh and frozen/thawed prepubertal mouse testicular tissues. The organotypic culture system will need to be further improved to increase the efficiency of in vitro spermatogenesis and allow a clinical application.

show abstract

Section: Discussionsupporting

confidence: 85%

Steroidogenesis and androgen/estrogen signaling pathways are altered inin vitromatured testicular tissues of prepubertal mice

Moutard

Goudin

Jaeger

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…Similar to other steroid receptors, it contains three domains: the amino-terminal domain (NTD), the DNA binding domain (DBD) and the ligand binding domain (LBD) [2]. Each domain has specific functions but communication as well as dimerization between different domains are required for full receptor activity [2][3][4][5].…”

Section: Introductionmentioning

confidence: 99%

The T850D Phosphomimetic Mutation in the Androgen Receptor Ligand Binding Domain Enhances Recruitment at Activation Function 2

Helsen

Nguyen

Vercruysse

et al. 2022

IJMS

Self Cite

View full text Add to dashboard Cite

Several key functions of the androgen receptor (AR) such as hormone recognition and co-regulator recruitment converge in the ligand binding domain (LBD). Loss- or gain-of-function of the AR contributes to pathologies such as the androgen insensitivity syndrome and prostate cancer. Here, we describe a gain-of-function mutation of the surface-exposed threonine at position 850, located at the amino-terminus of Helix 10 (H10) in the AR LBD. Since T850 phosphorylation was reported to affect AR function, we created the phosphomimetic mutation T850D. The AR T850D variant has a 1.5- to 2-fold increased transcriptional activity with no effect on ligand affinity. In the androgen responsive LNCaP cell line grown in medium with low androgen levels, we observed a growth advantage for cells in which the endogenous AR was replaced by AR T850D. Despite the distance to the AF2 site, the AR T850D LBD displayed an increased affinity for coactivator peptides as well as the 23FQNLF27 motif of AR itself. Molecular Dynamics simulations confirm allosteric transmission of the T850D mutation towards the AF2 site via extended hydrogen bond formation between coactivator peptide and AF2 site. This mechanistic study thus confirms the gain-of-function character of T850D and T850 phosphorylation for AR activity and reveals details of the allosteric communications within the LBD.

show abstract

“…Additionally, the AR defects caused by the V770A mutation could only be explained in the case of the GR‐like LBD dimer, further confirming the presence of a transitional or intermediate state between the AR LBD homodimer and the GR‐like LBD homodimer. Of note, the W751R mutation that demonstrated a disruptive effect on AR activity in mice (El Kharraz et al, 2021), destabilizes AR LBD dimerization in both models reflective of its central location in H5 of the LBD dimerization interface, which is the lynch pin in the middle of two transitional states.…”

Section: Discussionmentioning

confidence: 98%

Study of novel androgen receptor V770 variant in androgen insensitivity syndrome patients reveals the transitional state of the androgen receptor ligand binding domain homodimer

et al. 2023

Self Cite

View full text Add to dashboard Cite

We report the discovery of the androgen receptor missense mutation V770D, that was found in two sisters suffering from complete androgen insensitivity. Experimental validation of AR V770 variants demonstrated that AR V770D was transcriptionally inactive due to the inability to dimerize and a reduced ligand binding affinity. The more conservative AR V770A variant showed a dimerization defect at low levels of DHT with a partial recovery of the transcriptional activity and of the receptor's ability to dimerize when increasing the DHT levels. With V770 located outside of the proposed LBD dimerization interface of the AR LBD homodimer crystal structure, the effects of the V770A mutation on AR dimerization were unexpected. We therefore explored whether the AR LBD dimerization interface would be better described by an alternative dimerization mode based on available human homodimeric LBD crystal structures of other nuclear receptors. Superposition of the monomeric AR LBD in the homodimeric crystal structures of GR, PR, ER, CAR, TRβ, and HNF‐4α showed that the GR‐like LBD dimer model was energetically the most stable. Moreover, V770 was a key energy residue in the GR‐like LBD dimer while it was not involved in the stabilization of the AR LBD homodimer according to the crystal structure. Additionally, the observation that 4 AIS mutations impacted the stability of the AR LBD dimer while 16 mutations affected the GR‐like LBD dimer, suggested that the AR LBD dimer crystal is a snapshot of a breathing AR LBD homodimer that can transition into the GR‐like LBD dimer model.

show abstract

The androgen receptor depends on ligand‐binding domain dimerization for transcriptional activation

Cited by 30 publications

References 63 publications

Steroidogenesis and androgen/estrogen signaling pathways are altered inin vitromatured testicular tissues of prepubertal mice

Steroidogenesis and androgen/estrogen signaling pathways are altered inin vitromatured testicular tissues of prepubertal mice

The T850D Phosphomimetic Mutation in the Androgen Receptor Ligand Binding Domain Enhances Recruitment at Activation Function 2

Study of novel androgen receptor V770 variant in androgen insensitivity syndrome patients reveals the transitional state of the androgen receptor ligand binding domain homodimer

Contact Info

Product

Resources

About