Phosphoinositides regulate force-independent interactions between talin, vinculin, and actin
- Max Planck Institute of Biochemistry, Germany
Abstract
The focal adhesion (FA) proteins talin and vinculin connect integrin to actomyosin networks, acting as the core mechanosensitive FA machinery. Both proteins bind to F-actin and each other, providing a foundation for network formation within FAs. However, the underlying mechanisms regulating their engagement remain unclear. Here, we performed in vitro reconstitution of talin-vinculin-actin assemblies using synthetic membrane systems. Neither talin nor vinculin alone recruit actin filaments to the membrane. In contrast, phosphoinositide-rich membranes recruit and activate talin, and the membrane-bound talin then activates vinculin. Together, they link actin to the membrane. Encapsulation of these components within vesicles reorganized actin into higher-order networks. Notably, these observations were made in the absence of applied force, whereby we infer that the initial assembly stage of FAs is force independent. Our findings demonstrate that the local membrane composition plays a key role in controlling the stepwise recruitment, activation, and engagement of proteins within FAs.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files. Source data files would be provided for Figures 2,3,4,6 and 7.
Article and author information
Author details
Charlotte F Kelley
Funding
European Molecular Biology Organization (EMBO Long-term Fellowship Award)
- Charlotte F Kelley
Alexander von Humboldt-Stiftung (Research Fellowship for Postdoctoral Researchers)
- Charlotte F Kelley
Horizon 2020 Framework Programme (Marie Sklodowska-Curie Action)
- Charlotte F Kelley
Boehringer Ingelheim Stiftung (Plus 3)
- Naoko Mizuno
H2020 European Research Council (FocAd)
- Naoko Mizuno
European Molecular Biology Organization (Young Investigator Award)
- Naoko Mizuno
Max-Planck-Gesellschaft
- Naoko Mizuno
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.
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Phosphoinositides regulate force-independent interactions between talin, vinculin, and actin
eLife 9:e56110.
https://doi.org/10.7554/eLife.56110
