Visualizing the mechanical activation of Src

Yingxiao Wang, Elliot L. Botvinick, Yihua Zhao, Michael W. Berns, Shunichi Usami, Roger Y. Tsien and Shu Chien ()
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Yingxiao Wang: Department of Bioengineering and the Whitaker Institute of Biomedical Engineering
Elliot L. Botvinick: Department of Bioengineering and the Whitaker Institute of Biomedical Engineering
Yihua Zhao: Department of Bioengineering and the Whitaker Institute of Biomedical Engineering
Michael W. Berns: Department of Bioengineering and the Whitaker Institute of Biomedical Engineering
Shunichi Usami: Department of Bioengineering and the Whitaker Institute of Biomedical Engineering
Roger Y. Tsien: University of California at San Diego
Shu Chien: Department of Bioengineering and the Whitaker Institute of Biomedical Engineering

Nature, 2005, vol. 434, issue 7036, 1040-1045

Abstract: Abstract The mechanical environment crucially influences many cell functions1. However, it remains largely mysterious how mechanical stimuli are transmitted into biochemical signals. Src is known to regulate the integrin–cytoskeleton interaction2, which is essential for the transduction of mechanical stimuli3,4,5. 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 umbilical 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 stimulation site with a speed (mean ± s.e.m.) of 18.1 ± 1.7 nm s-1. This force-induced directional and long-range activation of Src was abolished by the disruption of actin filaments or microtubules. Our reporter has thus made it possible to monitor mechanotransduction in live cells with spatio-temporal characterization. We find that the transmission of mechanically induced Src activation is a dynamic process that directs signals via the cytoskeleton to spatial destinations.

Date: 2005
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DOI: 10.1038/nature03469

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