FORCEBIND: Mechanochemical Regulation Of Focal And Fibrillar Adhesion Proteins

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Cells sense and adapt to mechanical forces imposed by their environment. Mechanical cues from the extracellular matrix are detected at focal adhesions, where they are converted into biochemical signals to elicit a cellular response. A primary force-transduction mechanism involves the mechanical and chemical regulation of protein-binding interactions. Understanding these processes from a molecular perspective requires evaluating how these proteins interact under force. However, protein biochemistry assays cannot apply force to molecules in bulk, while classic single-molecule techniques are generally limited to using high forces, which precludes the study of mechanosensing proteins given the low forces involved at cell-matrix adhesions. Our objective is to implement novel single-molecule approaches to unravel the molecular mechanisms underpinning mechanotransduction processes at cellular adhesions. To that aim, we will first develop a new single-molecule instrument combining fluorescence with ultra-stable magnetic tweezers, allowing us to capture binding events in proteins under physiologically relevant forces and timescales. This new technology will enable us to examine how key mechanosensing proteins respond conformationally to force and how these forces regulate the binding interactions and post-translational modifications that underpin their function. Specifically, we will determine how phosphorylation regulates protein nanomechanics, using the focal adhesion kinase as a model system, and delve into the elusive mechanosensing function of tensin, the master regulator of fibrillar adhesions. We anticipate that these efforts will unveil how protein dynamics, interactions, and chemistry are finely regulated by mechanical forces, identifying the key molecular mechanisms used by cells to convert mechanical cues into biochemical responses, of critical relevance for multiple biological fields, from development to cancer.