Mechanotransduction in vivo by repeated talin stretch-relaxation events depends upon vinculin.
Mechanotransduction is a critical function for cells, in terms of cell viability, shaping of tissues, and cellular behavior. In vitro, cellular level forces can stretch adhesion proteins that link extracellular matrix to the actin cytoskeleton exposing hidden binding sites. However, there is no evidence that in vivo forces produce significant in vivo stretching to cause domain unfolding. We now report that the adhesion protein, talin, is repeatedly stretched by 100-350 nm in vivo by myosin contraction of actin filaments. Using a functional EGFP-N-Talin1-C-mCherry to measure the length of single talin molecules, we observed that the C-terminal mCherry was normally displaced in the direction of actin flow by 90 to >250 nm from N-EGFP but only by 50-60 nm (talin's length in vitro) after myosin inhibition. Individual talin molecules transiently stretched and relaxed. Peripheral, multimolecular adhesions had green outside and red proximal edges. They also exhibited transient, myosin-dependent stretching of 50-350 nm for 6-16 s; however, expression of the talin-binding head of vinculin increased stretching to about 400 nm and suppressed dynamics. We suggest that rearward moving actin filaments bind, stretch, and release talin in multiple, stochastic stick-slip cycles and that multiple vinculin binding and release cycles integrate pulling on matrices into biochemical signals.
Research Centre of Excellence in Mechanobiology, National University of Singapore, Singapore. felix@margadant.ch
Felix
F
Margadant
Li Li
LL
Chew
Xian
X
Hu
Hanry
H
Yu
Neil
N
Bate
Xian
X
Zhang
Michael
M
Sheetz
D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D002522:Chlorocebus aethiops; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000378:metabolism; D040542:Mechanotransduction, Cellular; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism; D016608:Talin / Q000096:biosynthesis* / Q000378:metabolism; D016596:Vinculin / Q000378:metabolism*
D011993:Recombinant Fusion Proteins; D016608:Talin; C485184:enhanced green fluorescent protein; D016596:Vinculin; D049452:Green Fluorescent Proteins