Mechano-coupling and regulation of contractility by the vinculin tail domain.
Vinculin binds to multiple focal adhesion and cytoskeletal proteins and has been implicated in transmitting mechanical forces between the actin cytoskeleton and integrins or cadherins. It remains unclear to what extent the mechano-coupling function of vinculin also involves signaling mechanisms. We report the effect of vinculin and its head and tail domains on force transfer across cell adhesions and the generation of contractile forces. The creep modulus and the adhesion forces of F9 mouse embryonic carcinoma cells (wild-type), vinculin knock-out cells (vinculin -/-), and vinculin -/- cells expressing either the vinculin head domain, tail domain, or full-length vinculin (rescue) were measured using magnetic tweezers on fibronectin-coated super-paramagnetic beads. Forces of up to 10 nN were applied to the beads. Vinculin -/- cells and tail cells showed a slightly higher incidence of bead detachment at large forces. Compared to wild-type, cell stiffness was reduced in vinculin -/- and head cells and was restored in tail and rescue cells. In all cell lines, the cell stiffness increased by a factor of 1.3 for each doubling in force. The power-law exponent of the creep modulus was force-independent and did not differ between cell lines. Importantly, cell tractions due to contractile forces were suppressed markedly in vinculin -/- and head cells, whereas tail cells generated tractions similar to the wild-type and rescue cells. These data demonstrate that vinculin contributes to the mechanical stability under large external forces by regulating contractile stress generation. Furthermore, the regulatory function resides in the tail domain of vinculin containing the paxillin-binding site.
Mierke
Claudia Tanja
CT
Center for Medical Physics and Technology, Department of Physics, Biophysics, University of Erlangen-Nuremberg, Erlangen, Germany. claudia.mierke@t-online.de
Kollmannsberger
Philip
P
Zitterbart
Daniel Paranhos
DP
Smith
James
J
Fabry
Ben
B
Goldmann
Wolfgang Heinrich
WH
D000818:Animals; D001696:Biomechanical Phenomena; D002449:Cell Aggregation; D045744:Cell Line, Tumor; D002465:Cell Movement; D040542:Mechanotransduction, Cellular; D051379:Mice; D008863:Microspheres; D009154:Mutation / Q000235:genetics; D017434:Protein Structure, Tertiary; D016596:Vinculin / Q000737:chemistry* / Q000378:metabolism*
D016596:Vinculin