BS42 Mechanical imprinting of the talin interaction with DLC1 regulates RHOA activity and cardiomyocyte remodelling
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Le résumé fourni par la source
Introduction During heart disease, the cardiac extracellular matrix (ECM) mechanically transforms, resulting in the stiffening of the ECM. Cardiomyocytes sense this ECM change, leading to phenotypic remodelling. A key part of the cardiomyocyte mechanosensing machinery is organised at the costamere; integrins and associated proteins, bridging the ECM and the sarcomere, facilitating force transmission. An integral component of the costamere is the mechanosensitive protein talin, with rod domains which unfold and refold in the presence and absence of physiological forces, exposing cryptic binding sites leading to force-dependent interactions with talin binding partners. While talin has been established as a mechanical sensor, the specifics of its mechanosensing pathway remain elusive. Methods Co-immunoprecipitation, fluorescence recovery after photobleaching, and fluorescence resonance energy transfer experiments were performed on neonate rat cardiomyocytes seeded on PDMS of stiffnesses corresponding to that of embryonic (1 kPa), healthy (6 kPa) and diseased (20–130 kPa) cardiac tissue. In situ proximity ligation was conducted on wildtype and MLP knockout mouse heart sections. The LOVTRAP optogenetics system was performed on the C2C12 mouse myoblast cell line, and fluorescence polarisation in vitro binding experiments used talin R7R8 domain, DLC1 and RIAM synthetic proteins expressed and purified from E. coli BL21. Adenoviral transduction and siRNA were used to overexpress and knockdown proteins, respectively. Results We identified three talin-interacting proteins, DLC1, RIAM and paxillin, which each preferentially bound to talin at specific ECM stiffnesses – paxillin preferred 1 kPa, DLC1 6 kPa and RIAM 20–130 kPa – and subsequently confirmed this in situ in healthy and diseased mouse hearts. The interactions between talin and DLC1 or RIAM were shown to be regulated through direct competition for binding at the cellular and molecular level, yielding Kd values of 4.9 µM and 9.2 µM for DLC1 and RIAM, respectively. These interactions were present even when talin was in a tension-free state during the immunoprecipitation, suggesting the competitive interactions might be further imprinted, i.e. stabilised through other mechanisms such as phosphorylation. Focal adhesion kinase activity was shown to regulate the interaction between talin and DLC1 or RIAM: inhibition of Src family kinases and FAK altered the talin interactions, promoting the talin-RIAM interaction over the talin-DLC1 at 6 kPa. Finally, we identified DLC1, as a major RhoGAP, regulated cardiomyocyte RhoA activity in a stiffness-dependent manner, with DLC1 knockdown resulting in cytoskeletal disruptions. Conclusions This work demonstrated a mechanism of imprinting mechanical information into the talin interactome to finetune RhoA activity, contributing to our knowledge of cardiomyocyte mechanosignalling which can help improve our understanding of cardiac health and disease.
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Le contrôle bibliographique ouvert
DOI retrouvé dans Crossref DOI retrouvé ; titre concordant.
- Titre Crossref
- BS42 Mechanical imprinting of the talin interaction with DLC1 regulates RHOA activity and cardiomyocyte remodelling
- Date Crossref
- 13/08/2025
- Éditeur
- BMJ Publishing Group Ltd and British Cardiovascular Society
- Type
- proceedings-article
Ce recoupement confirme des métadonnées liées au DOI. Il ne confirme ni la méthode ni les conclusions de l’étude, et il ne compte pas comme une seconde source scientifique indépendante.
Où se fait cette recherche
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Centre National de la Recherche Scientifique pays non établi dans la noticeOrganisme public
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Université de Bordeaux pays non établi dans la noticeUniversité ou école supérieure
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Queen Mary University of London pays non établi dans la noticeUniversité ou école supérieure
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Equilibrium Research pays non établi dans la noticeStructure de recherche
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Chimie et Biologie des Membranes et des Nanoobjects pays non établi dans la noticeStructure de recherche
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Institut Polytechnique de Bordeaux pays non établi dans la noticeUniversité ou école supérieure
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University of Liverpool Department of Biochemistry pays non établi dans la noticeUniversité ou école supérieure
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University of Birmingham Institute of Cardiovascular Sciences pays non établi dans la noticeUniversité ou école supérieure
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British Heart Foundation pays non établi dans la noticeOrganisation à but non lucratif
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University of Oxford Radcliffe Department of Medicine and British Heart Foundation Centre of Research Excellence Oxford pays non établi dans la noticeUniversité ou école supérieure
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Azienda Ospedaliera Universitaria Integrata Verona pays non établi dans la noticeÉtablissement de santé
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Imperial College London National Heart and Lung Institute pays non établi dans la noticeUniversité ou école supérieure
Centre National de la Recherche Scientifique, Université de Bordeaux et Queen Mary University of London, avec 9 autres affiliations.
Une affiliation ne permet pas de déduire la nationalité d’un auteur.