p66^Shc Couples Mechanical Signals to RhoA through Focal Adhesion Kinase-Dependent Recruitment of p115-RhoGEF and GEF-H1

Abstract

Tissue cells respond to changes in tensional forces with proliferation or death through the control of RhoA. However, the response coupling mechanisms that link force with RhoA activation are poorly understood. We found that tension applied to fibronectin-coated microbeads caused recruitment of all three isoforms of the Shc adapter (p66^Shc, p52^Shc, and p46^Shc) to adhesion complexes. The Shc PTB domain was necessary and sufficient for this recruitment, and screening studies revealed the direct interactions with the FERM domain of focal adhesion kinase (FAK) that were required for Shc translocation to adhesion complexes. The FAK/p66^Shc complex specifically bound and activated the Rho guanyl exchange factors (GEFs) p115-RhoGEF and GEF-H1, leading to tension-induced RhoA activation. In contrast, the FAK/p52^Shc complex bound SOS1 but not the Rho GEFs to mediate tension-induced Ras activation. Nuclear translocation and activation of the YAP/TAZ transcription factors on firm substrates required the FAK/p66^Shc/Rho GEF complex, and both proliferation on firm substrates and anoikis in suspension required signaling through p66^Shc and its associated Rho GEFs. These studies reveal the binary and exclusive assignment of p66^Shc and p52^Shc to tension-induced Rho or Ras signals, respectively, and suggest an integrated role for the two Shc isoforms in coordinating the cellular response to mechanical stimuli.

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ACKNOWLEDGMENTS

We thank Manasvini Ammanamanchi and Melanie Maurer for their assistance in mechanical testing of the hydrogels.

We acknowledge the University of Texas Southwestern for institutional support for the Proteomics Core Facility.