Agonist-controlled competition of RAR and VDR nuclear receptors for heterodimerization with RXR is manifested in their DNA binding

Rehó, Bálint; Fadel, Lina; Brázda, Péter; Benziane, Anass; Hegedüs, Éva; Sen, Pialy; Gadella, Theodorus; Tóth, Katalin; Nagy, László; Vámosi, György

doi:10.1016/j.jbc.2023.102896

Cited by 5 publications

(8 citation statements)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…According to the current consensus models for how nuclear receptor interaction networks operate, specific T2NRs members compete for a limited pool of the “core” partner RXR in cells, thus establishing a competitive regulatory network (Wood, 2008; Yoshikawa et al , 2003; Chan & Wells, 2009; Fadel et al , 2020; Wang et al , 2005; Rehó et al , 2023). However, previous in vivo studies were carried out with overexpressed proteins, limiting their ability to accurately address this model of competition (Fadel 2019, Reho 2023).…”

Section: Discussionmentioning

confidence: 99%

“…Whether such in vitro systems would capture the complexity and competitive dynamics at play in live cells has remained an unresolved issue. Moreover, to date, in vivo studies of competitive heterodimerization networks have not examined TFs at endogenous expression levels and thus may not accurately recapitulate their dynamic interactions with each other and with chromatin (Fadel et al, 2020;Grinberg et al, 2004) or do not account for expression levels of relevant TFs in individual cells due to the population averaging nature of most such studies (Rehó et al, 2023) (Reho 2023). To overcome these potential shortcomings, we employed fast singlemolecule tracking (fSMT) (Boka et al, 2021;Dahal et al, 2023;Elf et al, 2007;Hansen et al, 2018Hansen et al, , 2017 and its newly developed complement, proximity-assisted photoactivation (PAPA-SMT) (Graham et al, 2022) to test the effects of varying the stoichiometry of a core TF (RXRα) and its partner TF (RARα).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Surprising Features of Nuclear Receptor Interaction Networks Revealed by Live Cell Single Molecule Imaging

Dahal,

Graham,

Dailey

et al. 2023

Preprint

View full text Add to dashboard Cite

Type 2 Nuclear Receptors (T2NRs) require heterodimerization with a common partner, the Retinoid X Receptor (RXR), to bind cognate DNA recognition sites in chromatin. Based on previous biochemical and over-expression studies, binding of T2NRs to chromatin is proposed to be regulated by competition for a limiting pool of the core RXR subunit. However, this mechanism has not yet been tested for endogenous proteins in live cells. Using single molecule tracking (SMT) and proximity-assisted photoactivation (PAPA), we monitored interactions between endogenously tagged retinoid X receptor (RXR) and retinoic acid receptor (RAR) in live cells. Unexpectedly, we find that higher expression of RAR, but not RXR increases heterodimerization and chromatin binding in U2OS cells. This surprising finding indicates the limiting factor is not RXR but likely its cadre of obligate dimer binding partners. SMT and PAPA thus provide a direct way to probe which components are functionally limiting within a complex TF interaction network providing new insights into mechanisms of gene regulation in vivo with implications for drug development targeting nuclear receptors.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Surprising Features of Nuclear Receptor Interaction Networks Revealed by Live Cell Single Molecule Imaging

Dahal,

Graham,

Dailey

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…Similarly, it was found that bile acids generated in response to diet and biotransformed by certain bacteria induce colonic Foxp3 + RORγt + T cells via their interaction with the vitamin D receptor (VDR), which is highly expressed in colonic IECs, DCs and Tregs ( 116 ). VDR and the principal receptor for bile acids, farnesoid X receptor (FXF), form heterodimers with RXRs or RARs to function as ligand-induced transcription factors in target genes, and interaction between nuclear signalling by vitamin D or bile acids and RA has been documented ( 132 ). Bile acids have been found to stimulate a RARE response in DCs via FXR-RAR interactions ( 133 ) and, conversely, RA has been reported to upregulate FXR pathway genes and to modulate hepatic homeostasis and lipid metabolism ( 134 ).…”

Section: Intestinal Microbiome and Allergic Diseasementioning

confidence: 99%

Intestinal factors promoting the development of RORγt+ cells and oral tolerance

López-Fandiño,

Molina,

Lozano-Ojalvo

2023

Front. Immunol.

View full text Add to dashboard Cite

The gastrointestinal tract has to harmonize the two seemingly opposite functions of fulfilling nutritional needs and avoiding the entry of pathogens, toxins and agents that can cause physical damage. This balance requires a constant adjustment of absorptive and defending functions by sensing environmental changes or noxious substances and initiating adaptive or protective mechanisms against them through a complex network of receptors integrated with the central nervous system that communicate with cells of the innate and adaptive immune system. Effective homeostatic processes at barrier sites take the responsibility for oral tolerance, which protects from adverse reactions to food that cause allergic diseases. During a very specific time interval in early life, the establishment of a stable microbiota in the large intestine is sufficient to prevent pathological events in adulthood towards a much larger bacterial community and provide tolerance towards diverse food antigens encountered later in life. The beneficial effects of the microbiome are mainly exerted by innate and adaptive cells that express the transcription factor RORγt, in whose generation, mediated by different bacterial metabolites, retinoic acid signalling plays a predominant role. In addition, recent investigations indicate that food antigens also contribute, analogously to microbial-derived signals, to educating innate immune cells and instructing the development and function of RORγt+ cells in the small intestine, complementing and expanding the tolerogenic effect of the microbiome in the colon. This review addresses the mechanisms through which microbiota-produced metabolites and dietary antigens maintain intestinal homeostasis, highlighting the complementarity and redundancy between their functions.

show abstract

“…Such interactions are routinely revealed in bulk in vitro samples by immunoprecipitation; however, to show that they occur in situ in the cell and could possibly be disrupted by a pharmacological agent requires sensitive microscopic approaches [ 38 ]. Fluorescence correlation spectroscopy (FCS) and fluorescence cross-correlation spectroscopy (FCCS) are advanced methods with potential single molecule sensitivity that have currently started to gain popularity due to their potential for sensitively assessing molecular mobility and dynamic interactions in situ in living cells [ 39 , 40 , 41 , 42 , 43 , 44 ]. Since alterations in molecular mobility and consequential changes in characteristic diffusion times are not highly sensitive to dimerization or low levels of aggregation, these associations are best monitored through the photon-count histogram (PCH), which is highly sensitive to oligomerization [ 45 , 46 ].…”

Section: Introductionmentioning

confidence: 99%

Disrupting EGFR–HER2 Transactivation by Pertuzumab in HER2-Positive Cancer: Quantitative Analysis Reveals EGFR Signal Input as Potential Predictor of Therapeutic Outcome

Ujlaky-Nagy,

Szöllősi,

Vereb

2024

IJMS

View full text Add to dashboard Cite

Pertuzumab (Perjeta®), a humanized antibody binding to the dimerization arm of HER2 (Human epidermal growth factor receptor-2), has failed as a monotherapy agent in HER2 overexpressing malignancies. Since the molecular interaction of HER2 with ligand-bound EGFR (epidermal growth factor receptor) has been implied in mitogenic signaling and malignant proliferation, we hypothesized that this interaction, rather than HER2 expression and oligomerization alone, could be a potential molecular target and predictor of the efficacy of pertuzumab treatment. Therefore, we investigated static and dynamic interactions between HER2 and EGFR molecules upon EGF stimulus in the presence and absence of pertuzumab in HER2+ EGFR+ SK-BR-3 breast tumor cells using Förster resonance energy transfer (FRET) microscopy and fluorescence correlation and cross-correlation spectroscopy (FCS/FCCS). The consequential activation of signaling and changes in cell proliferation were measured by Western blotting and MTT assay. The autocorrelation functions of HER2 diffusion were best fitted by a three-component model corrected for triplet formation, and among these components the slowly diffusing membrane component revealed aggregation induced by EGFR ligand binding, as evidenced by photon-counting histograms and co-diffusing fractions. This aggregation has efficiently been prevented by pertuzumab treatment, which also inhibited the post-stimulus interaction of EGFR and HER2, as monitored by changes in FRET efficiency. Overall, the data demonstrated that pertuzumab, by hindering post-stimulus interaction between EGFR and HER2, inhibits EGFR-evoked HER2 aggregation and phosphorylation and leads to a dose-dependent decrease in cell proliferation, particularly when higher amounts of EGF are present. Consequently, we propose that EGFR expression on HER2-positive tumors could be taken into consideration as a potential biomarker when predicting the outcome of pertuzumab treatment.

show abstract

Agonist-controlled competition of RAR and VDR nuclear receptors for heterodimerization with RXR is manifested in their DNA binding

Cited by 5 publications

References 55 publications

Surprising Features of Nuclear Receptor Interaction Networks Revealed by Live Cell Single Molecule Imaging

Surprising Features of Nuclear Receptor Interaction Networks Revealed by Live Cell Single Molecule Imaging

Intestinal factors promoting the development of RORγt+ cells and oral tolerance

Disrupting EGFR–HER2 Transactivation by Pertuzumab in HER2-Positive Cancer: Quantitative Analysis Reveals EGFR Signal Input as Potential Predictor of Therapeutic Outcome

Contact Info

Product

Resources

About