In-depth analysis of the mode of action of resveratrol: genome-wide characterization of G-quadruplex binding properties
Abstract Background Resveratrol (RSV) is one of the most studied and used biomolecules, for which many pharmacological effects targeting multiple tissues have been described. However, a common underlying mechanism driving its full pharmacological activity has not been described in detail. G-quadrupl...
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2025-06-01
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| Series: | Cellular & Molecular Biology Letters |
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| Online Access: | https://doi.org/10.1186/s11658-025-00747-1 |
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| author | Ana Soriano-Lerma Victoria Sánchez-Martín Javier Murciano-Calles Matilde Ortiz-González María J. Tello-López Virginia Pérez-Carrasco Ángel Linde-Rodríguez Inmaculada Ramírez-Macías Irene Gómez-Pìnto Inmaculada López-Aliaga Miguel Soriano Jose A. Garcia Salcedo |
| author_facet | Ana Soriano-Lerma Victoria Sánchez-Martín Javier Murciano-Calles Matilde Ortiz-González María J. Tello-López Virginia Pérez-Carrasco Ángel Linde-Rodríguez Inmaculada Ramírez-Macías Irene Gómez-Pìnto Inmaculada López-Aliaga Miguel Soriano Jose A. Garcia Salcedo |
| author_sort | Ana Soriano-Lerma |
| collection | DOAJ |
| description | Abstract Background Resveratrol (RSV) is one of the most studied and used biomolecules, for which many pharmacological effects targeting multiple tissues have been described. However, a common underlying mechanism driving its full pharmacological activity has not been described in detail. G-quadruplexes (G4s) are non-canonical nucleic acid structures found in regulatory genomic locations and involved in controlling gene transcription, telomere maintenance, or genome stability, among others. This study provides a genome-wide characterization of RSV G4-binding properties, explaining its multi-target traits. Methods Immunofluorescence assays using a nucleolar and a G4-specific antibody were used to characterize RSV cellular effects on the nucleolus and G4 stabilization. DNA damage and cell cycle analyses were performed via western blot and flow cytometry. Breaks lLbeling In Situ and Sequencing (BLISS) was used to map double strand breaks (DSB) in response to treatment, and identify G4s targeted by RSV. mRNA sequencing was used to identify changes at the transcriptional level upon treatment and relate them to a direct targeting of G4s. Biophysical assays (circular dichroism, ultraviolet–visible [UV–Vis] titration, differential scanning calorimetry, and nuclear magnetic resonance) were used to characterize RSV–G4 interactions. Lastly, luciferase-based transcription assays were performed to confirm RSV–G4 interaction in vitro and its direct influence on gene expression. Results In a cellular context, RSV treatment showed classic G4-ligand effects, such as nucleolar disassembly, inhibition of RNA polymerase I, DNA damage, and cell cycle arrest. RSV was shown to stabilize cellular G4s, which accumulated around double strand breaks in the promoters of differentially expressed genes. Upon treatment, G4 stabilization triggered DNA damage and controlled gene expression. The interaction between RSV and target G4s was confirmed in vitro by biophysical assays and through luciferase-based transcription assays. Conclusions A G4-dependent mode of action was demonstrated as the main mechanism underlying RSV pleiotropic effects, along with the identification of target genes and G4s. This in-depth analysis of the mode of action of RSV will be helpful to improve its therapeutic potential in a wide variety of health scenarios. Graphical Abstract |
| format | Article |
| id | doaj-art-814ab4caefe24a168fa91361be236eb5 |
| institution | OA Journals |
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| publishDate | 2025-06-01 |
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| spelling | doaj-art-814ab4caefe24a168fa91361be236eb52025-08-20T02:37:33ZengBMCCellular & Molecular Biology Letters1689-13922025-06-0130112510.1186/s11658-025-00747-1In-depth analysis of the mode of action of resveratrol: genome-wide characterization of G-quadruplex binding propertiesAna Soriano-Lerma0Victoria Sánchez-Martín1Javier Murciano-Calles2Matilde Ortiz-González3María J. Tello-López4Virginia Pérez-Carrasco5Ángel Linde-Rodríguez6Inmaculada Ramírez-Macías7Irene Gómez-Pìnto8Inmaculada López-Aliaga9Miguel Soriano10Jose A. Garcia Salcedo11GENYO, Centre for Genomics and Oncological Research: Pfizer/University of Granada/Andalusian Regional GovernmentDepartment of Biochemistry, Molecular Biology III and Immunology, University of GranadaDepartment of Physical Chemistry, Unit of Excellence for Chemistry Applied to Biomedicine and the Environment, and Institute of Biotechnology, University of GranadaCenter for Intensive Mediterranean Agrosystems and Agri-Food Biotechnology (CIAIMBITAL), University of AlmeriaGENYO, Centre for Genomics and Oncological Research: Pfizer/University of Granada/Andalusian Regional GovernmentGENYO, Centre for Genomics and Oncological Research: Pfizer/University of Granada/Andalusian Regional GovernmentGENYO, Centre for Genomics and Oncological Research: Pfizer/University of Granada/Andalusian Regional GovernmentDepartment of Parasitology, University of GranadaInstituto de Química Física ‘Blas Cabrera’, (IQF-CSIC)Department of Physiology (Faculty of Pharmacy), Institute of Nutrition and Food Technology “José Mataix”, University of GranadaCenter for Intensive Mediterranean Agrosystems and Agri-Food Biotechnology (CIAIMBITAL), University of AlmeriaGENYO, Centre for Genomics and Oncological Research: Pfizer/University of Granada/Andalusian Regional GovernmentAbstract Background Resveratrol (RSV) is one of the most studied and used biomolecules, for which many pharmacological effects targeting multiple tissues have been described. However, a common underlying mechanism driving its full pharmacological activity has not been described in detail. G-quadruplexes (G4s) are non-canonical nucleic acid structures found in regulatory genomic locations and involved in controlling gene transcription, telomere maintenance, or genome stability, among others. This study provides a genome-wide characterization of RSV G4-binding properties, explaining its multi-target traits. Methods Immunofluorescence assays using a nucleolar and a G4-specific antibody were used to characterize RSV cellular effects on the nucleolus and G4 stabilization. DNA damage and cell cycle analyses were performed via western blot and flow cytometry. Breaks lLbeling In Situ and Sequencing (BLISS) was used to map double strand breaks (DSB) in response to treatment, and identify G4s targeted by RSV. mRNA sequencing was used to identify changes at the transcriptional level upon treatment and relate them to a direct targeting of G4s. Biophysical assays (circular dichroism, ultraviolet–visible [UV–Vis] titration, differential scanning calorimetry, and nuclear magnetic resonance) were used to characterize RSV–G4 interactions. Lastly, luciferase-based transcription assays were performed to confirm RSV–G4 interaction in vitro and its direct influence on gene expression. Results In a cellular context, RSV treatment showed classic G4-ligand effects, such as nucleolar disassembly, inhibition of RNA polymerase I, DNA damage, and cell cycle arrest. RSV was shown to stabilize cellular G4s, which accumulated around double strand breaks in the promoters of differentially expressed genes. Upon treatment, G4 stabilization triggered DNA damage and controlled gene expression. The interaction between RSV and target G4s was confirmed in vitro by biophysical assays and through luciferase-based transcription assays. Conclusions A G4-dependent mode of action was demonstrated as the main mechanism underlying RSV pleiotropic effects, along with the identification of target genes and G4s. This in-depth analysis of the mode of action of RSV will be helpful to improve its therapeutic potential in a wide variety of health scenarios. Graphical Abstracthttps://doi.org/10.1186/s11658-025-00747-1Small moleculeDNA bindingG-quadruplexResveratrolSecondary DNA structures |
| spellingShingle | Ana Soriano-Lerma Victoria Sánchez-Martín Javier Murciano-Calles Matilde Ortiz-González María J. Tello-López Virginia Pérez-Carrasco Ángel Linde-Rodríguez Inmaculada Ramírez-Macías Irene Gómez-Pìnto Inmaculada López-Aliaga Miguel Soriano Jose A. Garcia Salcedo In-depth analysis of the mode of action of resveratrol: genome-wide characterization of G-quadruplex binding properties Cellular & Molecular Biology Letters Small molecule DNA binding G-quadruplex Resveratrol Secondary DNA structures |
| title | In-depth analysis of the mode of action of resveratrol: genome-wide characterization of G-quadruplex binding properties |
| title_full | In-depth analysis of the mode of action of resveratrol: genome-wide characterization of G-quadruplex binding properties |
| title_fullStr | In-depth analysis of the mode of action of resveratrol: genome-wide characterization of G-quadruplex binding properties |
| title_full_unstemmed | In-depth analysis of the mode of action of resveratrol: genome-wide characterization of G-quadruplex binding properties |
| title_short | In-depth analysis of the mode of action of resveratrol: genome-wide characterization of G-quadruplex binding properties |
| title_sort | in depth analysis of the mode of action of resveratrol genome wide characterization of g quadruplex binding properties |
| topic | Small molecule DNA binding G-quadruplex Resveratrol Secondary DNA structures |
| url | https://doi.org/10.1186/s11658-025-00747-1 |
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