title: Distinct focal adhesion protein modules control different aspects of mechanotransduction.
abstract: Focal adhesions (FAs) are macromolecular complexes that regulate cell adhesion and mechanotransduction. By performing fluorescence recovery after photobleaching (FRAP) and fluorescence loss after photoactivation (FLAP) experiments, we found that the mobility of core FA proteins correlates with their function. Structural proteins such as tensin, talin and vinculin are significantly less mobile in FAs than signaling proteins such as FAK (also known as PTK2) and paxillin. The mobilities of the structural proteins are directly influenced by substrate stiffness, suggesting that they are involved in sensing the rigidity of the extracellular environment. The turnover rates of FAK and paxillin, as well as kindlin2 (also known as FERMT2), are not influenced by substrate stiffness. By using specific Src and FAK inhibitors, we reveal that force-sensing by vinculin occurs independently of FAK and paxillin phosphorylation. However, their phosphorylation is required for downstream Rac1-driven cellular processes, such as protrusion and cell migration. Overall, we show that the FA is composed of different functional modules that separately control mechanosensing and the cellular mechano-response.
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: Ben
authors.identifier: 0000-0002-4231-9314
authors.initials: B
authors.lastname: Stutchbury
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: Paul
authors.identifier: 0000-0003-3098-674X
authors.initials: P
authors.lastname: Atherton
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: Ricky
authors.identifier: 0000-0003-0274-9827
authors.initials: R
authors.lastname: Tsang
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester M13 9PT, England, UK.
authors.forename: De-Yao
authors.identifier: 0000-0001-6431-4680
authors.initials: DY
authors.lastname: Wang
authors.affiliation: Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, Manchester M13 9PT, England, UK christoph.ballestrem@manchester.ac.uk.
authors.forename: Christoph
authors.identifier: 0000-0002-5375-7985
authors.initials: C
authors.lastname: Ballestrem
mesh_terms: D000818:Animals; D002465:Cell Movement; D005109:Extracellular Matrix / Q000378:metabolism; D036681:Fluorescence Recovery After Photobleaching; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D022001:Focal Adhesions / Q000378:metabolism*; D040542:Mechanotransduction, Cellular; D051379:Mice; D008954:Models, Biological; D041681:NIH 3T3 Cells; D051419:Paxillin / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D021381:Protein Transport; D011554:Pseudopodia / Q000378:metabolism; D015398:Signal Transduction; D016596:Vinculin / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism
chemical_list: D051419:Paxillin; D016596:Vinculin; D019000:Phosphotyrosine; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019061:src-Family Kinases
keywords: FAK; Focal adhesion; Mechanotransduction; Paxillin; Talin; Vinculin