Abstract
Enabled/vasodilator-stimulated phosphoprotein (Ena/VASP) proteins are established regulators of actin-based motility, platelet aggregation, and growth cone guidance. However, the molecular mechanisms involved essentially remain elusive. Here we report on a novel mechanism of VASP action, namely the regulation of tensile strength, contractility, and rigidity of the actin cytoskeleton. Compared to wild-type cells fibroblasts derived from VASP-deficient mice have thicker and more stable actin stress fibres. Furthermore focal adhesions are enlarged, myosin light chain phosphorylation is increased, and the rigidity of the filament-supported plasma membrane is elevated about three- to fourfold, as is evident from atomic force microscopy. Moreover, fibronectin-coated beads adhere stronger to the surface of VASP-deficient cells. The resistance of these beads to mechanical displacement by laser tweezers is dramatically increased in an F-actin-dependent mode. Cytoskeletal stabilization coincides with slower cell adhesion and detachment, while overall adhesion is increased. Interestingly, many of these effects observed in VASP (−/−) cells are recapitulated in VASP-overexpressing cells, hinting towards a balanced stoichiometry necessary for appropriate VASP function. Taken together, our results suggest that VASP regulates surface protrusion formation and cell adhesion through modulation of the mechanical properties of the actin cytoskeleton.
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Acknowledgments
The authors thank Elke Baumeister, Agnes Weth, Elke Butt, and Stepan Gambaryan for help and reagents, and Ulrich Walter for continuous support and discussions. This work was supported by the Deutsche Forschungsgemeinschaft (SFB 487 B4 & B5, SFB 355 B4 & C8, and Re1011/3-2).
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Annette B. Galler, Maísa I. García Arguinzonis these authors contributed equally to this work
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Galler, A.B., García Arguinzonis, M.I., Baumgartner, W. et al. VASP-dependent regulation of actin cytoskeleton rigidity, cell adhesion, and detachment. Histochem Cell Biol 125, 457–474 (2006). https://doi.org/10.1007/s00418-005-0091-z
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DOI: https://doi.org/10.1007/s00418-005-0091-z