Visualizing the mechanical activation of Src

  • Immunotherapeutic D
  • Immunol R
  • Wang Y
  • et al.
ISSN: 1476-4687
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Abstract

The mechanical environment crucially influences many cell functions1 .However, it remains largelymysterious howmechani- cal stimuli are transmitted into biochemical signals. Src is known to regulate the integrin–cytoskeleton interaction2 ,which is essential for the transduction of mechanical stimuli3–5 . Using fluorescent resonance energy transfer (FRET), here we develop a genetically encoded Src reporter that enables the imaging and quantification of spatio-temporal activation of Src in live cells. We introduced a local mechanical stimulation to human umbili- cal vein endothelial cells (HUVECs) by applying laser-tweezer traction on fibronectin-coated beads adhering to the cells. Using the Src reporter, we observed a rapid distal Src activation and a slower directional wave propagation of Src activation along the plasma membrane. This wave propagated away from the stimu- lation site with a speed (mean 6 s.e.m.) of 18.1 6 1.7nms21. This force-induced directional and long-range activation of Src was abolished by the disruption of actin filaments or micro- tubules. Our reporter has thus made it possible to monitor mechanotransduction in live cells with spatio-temporal charac- terization.We find that the transmission of mechanically induced Src activation is a dynamic process that directs signals via the cytoskeleton to spatial destinations.

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Immunotherapeutic, D. M., Immunol, R., Wang, Y. et al, Botvinick, E. L., Zhao, Y., & Berns, M. W. (2005). Visualizing the mechanical activation of Src. Nature, 434(7034), 772–7.

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