Mechanical forces are involved in many physiological processes. While most of the current work in mechanotransduction has focused on the function of focal adhesion proteins and the actin cytoskeleton, the mechanical role of intracellular and nuclear proteins remains rather elusive. In particular, the role of the LINC (Linker of Nucleoskeleton and Cytoskeleton) complex an ensemble of proteins that links the cytoskeleton network with the nucleus has received comparatively much less attention. This project aims at studying the nanomechanical properties of the individual proteins forming the LINC complex at the single molecule level, using single molecule force-clamp spectroscopy AFM and Magnetic Tweezers. In particular, we will investigate, for each protein, the reversibility, kinetics and effect of post-translational modifications in their individual mechanical unfolding/refolding trajectories, of key importance in proteins that are thought to play an important role as mechanosensors. Finally, we will compare the in-vitro results obtained at the single molecule level with the behaviour of the LINC proteins in live cells under compressing and stretching forces. These experiments will shed new light onto the mechanisms of force transmission to the cell nucleus on biologically relevant timescales.
Funding: 4 years, funded through a Leverhulme Trust Grant (stipend in line with MRC rates)
Subject Areas: Physical Sciences, single molecule biophysics, Biological sciences, physical chemistry, proteins, magnetic tweezers, AFM.
For more information, please contact Professor Sergi Garcia-Manyes at [email protected]
Beedle, A. E*., Mora, M*., Davis, C., Snijders, A., Stirnemann, G., Garcia-Manyes, S. «Forcing the reversibility of a mechanochemical reaction», Nature Communications (2018), 9, 3155.
Perales-Calvo, J., Giganti, D., Stirnemann, G., Garcia-Manyes, S. «The force-dependent mechanisms of DnaK-mediated mechanical folding», Science Advances (2018) 4, 2.
Garcia-Manyes, S., Beedle, A.E.M. «Steering chemical reactions with force» Nature Reviews Chemistry (2017).
Elosegui-Artola, A., Andreu, I., Beedle, A.E.M., Lezamiz, A., Uroz, M., Kosmalska, A., Oria, R., Kechagia, J., Rico-Lastres, P., Le Roux, A., Shanahan, C., Trepat, X., Navajas, D., Garcia-Manyes, S., Roca-Cusachs, P. «Force triggers YAP entry by mechanically regulating transport across nuclear pores» Cell (2017), 171, 1-14.
Beedle, A. E., Mora, M., Lynham, S., Stirnemann, G., Garcia-Manyes, S. «Tailoring protein nanomechanics with chemical reactivity» Nature Communications (2017), 8, 15658.
Beedle, A. E.; Lynham, S.; Garcia-Manyes, S. « Protein S-sulfenylation is a fleeting molecular switch that regulates non-enzymatic oxidative folding » Nature Communications (2016), 7, 12490.