Allosteric activation of vinculin by talin

Florian Franz; Rafael Tapia Rojo; Sabina  Winograd-Katz; Rajaa Boujemaa-Paterski; wending li; tamar Unger; shira albeck; camilo aponte-santamaria; Sergi Garcia-Manyes; Ohad Medalia; Benjamin geiger; frauke grater

doi:10.1038/s41467-023-39646-4

Allosteric activation of vinculin by talin

Florian Franz, Rafael Tapia Rojo^*, Sabina Winograd-Katz, Rajaa Boujemaa-Paterski, wending li, tamar Unger, shira albeck, camilo aponte-santamaria, Sergi Garcia-Manyes, Ohad Medalia^*, Benjamin geiger^*, frauke grater^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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Abstract

The talin-vinculin axis is a key mechanosensing component of cellular focal adhesions. How talin and vinculin respond to forces and regulate one another remains unclear. By combining single-molecule magnetic tweezers experiments, Molecular Dynamics simulations, actin-bundling assays, and adhesion assembly experiments in live cells, we here describe a two-ways allosteric network within vinculin as a regulator of the talin-vinculin interaction. We directly observe a maturation process of vinculin upon talin binding, which reinforces the binding to talin at a rate of 0.03 s−1. This allosteric transition can compete with force-induced dissociation of vinculin from talin only at forces up to 10 pN. Mimicking the allosteric activation by mutation yields a vinculin molecule that bundles actin and localizes to focal adhesions in a force-independent manner. Hence, the allosteric switch confines talin-vinculin interactions and focal adhesion build-up to intermediate force levels. The ‘allosteric vinculin mutant’ is a valuable molecular tool to further dissect the mechanical and biochemical signalling circuits at focal adhesions and elsewhere.

Original language	English
Article number	4311
Journal	Nature Communications
Volume	14
Issue number	4311
DOIs	https://doi.org/10.1038/s41467-023-39646-4
Publication status	Published - 18 Jul 2023

Keywords

Vinculin
Talin
Mechanotransduction

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title = "Allosteric activation of vinculin by talin",

abstract = "The talin-vinculin axis is a key mechanosensing component of cellular focal adhesions. How talin and vinculin respond to forces and regulate one another remains unclear. By combining single-molecule magnetic tweezers experiments, Molecular Dynamics simulations, actin-bundling assays, and adhesion assembly experiments in live cells, we here describe a two-ways allosteric network within vinculin as a regulator of the talin-vinculin interaction. We directly observe a maturation process of vinculin upon talin binding, which reinforces the binding to talin at a rate of 0.03 s−1. This allosteric transition can compete with force-induced dissociation of vinculin from talin only at forces up to 10 pN. Mimicking the allosteric activation by mutation yields a vinculin molecule that bundles actin and localizes to focal adhesions in a force-independent manner. Hence, the allosteric switch confines talin-vinculin interactions and focal adhesion build-up to intermediate force levels. The {\textquoteleft}allosteric vinculin mutant{\textquoteright} is a valuable molecular tool to further dissect the mechanical and biochemical signalling circuits at focal adhesions and elsewhere.",

keywords = "Vinculin, Talin, Mechanotransduction",

author = "Florian Franz and {Tapia Rojo}, Rafael and Sabina Winograd-Katz and Rajaa Boujemaa-Paterski and wending li and tamar Unger and shira albeck and camilo aponte-santamaria and Sergi Garcia-Manyes and Ohad Medalia and Benjamin geiger and frauke grater",

note = "Funding Information: F.G. and B.G. thank the Klaus Tschira Foundation for generous financial support. O.M. and R.B.P. thank the ZMB and ScopeM microscopy centers at UZH and ETH, respectively, for their support and assistance. The study was also supported by the Minerva Center on “Aging, from Physical Materials to Human Tissues” to B.G.; the Schweizerischer Nationalfonds zur F{\"o}rderung der Wissenschaftlichen Forschung to O.M. (SNSF 310030_207453); the HGS MathComp graduate school of Heidelberg University, the Max Planck School Matter to Life supported by the German Federal Ministry of Education and Research (BMBF) in collaboration with the Max Planck Society, the Flagship Initiative funded by the Federal Ministry of Education and Research (BMBF) and the Ministry of Science Baden-W{\"u}rttemberg within the framework of the Excellence Strategy of the Federal and State Governments of Germany, the state of Baden-W{\"u}rttemberg through bwHPC and the German Research Foundation (DFG) through grant INST 35/1134-1 FUGG to F.G. This work was supported in part by the Francis Crick Institute which receives its core funding from Cancer Research U.K. (FC001002), the U.K. Medical Research Council (FC001002), and the Wellcome Trust (FC001002). R.T.-R. is recipient of a King{\textquoteright}s Prize Fellowship. This work was supported by the European Commission (Mechanocontrol, Grant Agreement 731957), BBSRC sLoLa (BB/V003518/1), Leverhulme Trust Research Leadership Award RL 2016-015, Wellcome Trust Investigator Award 212218/Z/18/Z and Royal Society Wolfson Fellowship RSWF/R3/183006 to S.G.M, and by the European Commission (RADICOL, 101002812) to F.G. Publisher Copyright: {\textcopyright} 2023, The Author(s).",

year = "2023",

month = jul,

day = "18",

doi = "10.1038/s41467-023-39646-4",

language = "English",

volume = "14",

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T1 - Allosteric activation of vinculin by talin

AU - Franz, Florian

AU - Tapia Rojo, Rafael

AU - Winograd-Katz, Sabina

AU - Boujemaa-Paterski, Rajaa

AU - li, wending

AU - Unger, tamar

AU - albeck, shira

AU - aponte-santamaria, camilo

AU - Garcia-Manyes, Sergi

AU - Medalia, Ohad

AU - geiger, Benjamin

AU - grater, frauke

N1 - Funding Information: F.G. and B.G. thank the Klaus Tschira Foundation for generous financial support. O.M. and R.B.P. thank the ZMB and ScopeM microscopy centers at UZH and ETH, respectively, for their support and assistance. The study was also supported by the Minerva Center on “Aging, from Physical Materials to Human Tissues” to B.G.; the Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung to O.M. (SNSF 310030_207453); the HGS MathComp graduate school of Heidelberg University, the Max Planck School Matter to Life supported by the German Federal Ministry of Education and Research (BMBF) in collaboration with the Max Planck Society, the Flagship Initiative funded by the Federal Ministry of Education and Research (BMBF) and the Ministry of Science Baden-Württemberg within the framework of the Excellence Strategy of the Federal and State Governments of Germany, the state of Baden-Württemberg through bwHPC and the German Research Foundation (DFG) through grant INST 35/1134-1 FUGG to F.G. This work was supported in part by the Francis Crick Institute which receives its core funding from Cancer Research U.K. (FC001002), the U.K. Medical Research Council (FC001002), and the Wellcome Trust (FC001002). R.T.-R. is recipient of a King’s Prize Fellowship. This work was supported by the European Commission (Mechanocontrol, Grant Agreement 731957), BBSRC sLoLa (BB/V003518/1), Leverhulme Trust Research Leadership Award RL 2016-015, Wellcome Trust Investigator Award 212218/Z/18/Z and Royal Society Wolfson Fellowship RSWF/R3/183006 to S.G.M, and by the European Commission (RADICOL, 101002812) to F.G. Publisher Copyright: © 2023, The Author(s).

PY - 2023/7/18

Y1 - 2023/7/18

N2 - The talin-vinculin axis is a key mechanosensing component of cellular focal adhesions. How talin and vinculin respond to forces and regulate one another remains unclear. By combining single-molecule magnetic tweezers experiments, Molecular Dynamics simulations, actin-bundling assays, and adhesion assembly experiments in live cells, we here describe a two-ways allosteric network within vinculin as a regulator of the talin-vinculin interaction. We directly observe a maturation process of vinculin upon talin binding, which reinforces the binding to talin at a rate of 0.03 s−1. This allosteric transition can compete with force-induced dissociation of vinculin from talin only at forces up to 10 pN. Mimicking the allosteric activation by mutation yields a vinculin molecule that bundles actin and localizes to focal adhesions in a force-independent manner. Hence, the allosteric switch confines talin-vinculin interactions and focal adhesion build-up to intermediate force levels. The ‘allosteric vinculin mutant’ is a valuable molecular tool to further dissect the mechanical and biochemical signalling circuits at focal adhesions and elsewhere.

AB - The talin-vinculin axis is a key mechanosensing component of cellular focal adhesions. How talin and vinculin respond to forces and regulate one another remains unclear. By combining single-molecule magnetic tweezers experiments, Molecular Dynamics simulations, actin-bundling assays, and adhesion assembly experiments in live cells, we here describe a two-ways allosteric network within vinculin as a regulator of the talin-vinculin interaction. We directly observe a maturation process of vinculin upon talin binding, which reinforces the binding to talin at a rate of 0.03 s−1. This allosteric transition can compete with force-induced dissociation of vinculin from talin only at forces up to 10 pN. Mimicking the allosteric activation by mutation yields a vinculin molecule that bundles actin and localizes to focal adhesions in a force-independent manner. Hence, the allosteric switch confines talin-vinculin interactions and focal adhesion build-up to intermediate force levels. The ‘allosteric vinculin mutant’ is a valuable molecular tool to further dissect the mechanical and biochemical signalling circuits at focal adhesions and elsewhere.

KW - Vinculin

KW - Talin

KW - Mechanotransduction

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U2 - 10.1038/s41467-023-39646-4

DO - 10.1038/s41467-023-39646-4

M3 - Article

SN - 2041-1723

VL - 14

JO - Nature Communications

JF - Nature Communications

IS - 4311

M1 - 4311

ER -

Allosteric activation of vinculin by talin

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