Thrombospondin induces RhoA inactivation through FAK-dependent signaling to stimulate focal adhesion disassembly

Anthony Wayne Orr, Manuel Antonio Pallero, Wencheng Xiong, Joanne E. Murphy-Ullrich

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Cells utilize dynamic interactions with the extracellular matrix to adapt to changing environmental conditions. Thrombospondin 1 (TSP1) induces focal adhesion disassembly and cell migration through a sequence (hep I) in its heparin-binding domain signaling through the calreticulin-low density lipoprotein receptor-related protein receptor complex. This involves the Gαi-dependent activation of ERK and phosphoinositide (PI) 3-kinase, both of which are required for focal adhesion disassembly. Focal adhesion kinase (FAK) regulates adhesion dynamics, acting in part by modulating RhoA activity, and FAK is implicated in ERK and PI 3-kinase activation. In this work, we sought to determine the role of FAK in TSP1-induced focal adhesion disassembly. TSP1/hep I does not stimulate focal adhesion disassembly in FAK knockout fibroblasts, whereas re-expressing FAK rescues responsiveness. Inhibiting FAK signaling through FBNK or FAK Y397F expression in endothelial cells also abrogates this response. TSP1/hep I stimulates a transient increase in FAK phosphorylation that requires calreticulin and Gαi, but not ERK or PI 3-kinase. Hep I does not activate ERK or PI 3-kinase in FAK knockout fibroblasts, suggesting activation occurs downstream of FAK. TSP1/hep I stimulates RhoA inactivation with kinetics corresponding to focal adhesion disassembly in a FAK, ERK, and PI 3-ldnase-dependent manner. Furthermore, hep I does not stimulate focal adhesion disassembly in cells expressing constitutively active RhoA, suggesting that RhoA inactivation is required for this response. This is the first work to illustrate a connection between FAK phosphorylation in response to a soluble factor and MhoA inactivation, as well as the first report of PI 3-kinase and ERK in FAK regulation of RhoA activity.

Original languageEnglish (US)
Pages (from-to)48983-48992
Number of pages10
JournalJournal of Biological Chemistry
Volume279
Issue number47
DOIs
StatePublished - Nov 19 2004

Fingerprint

Thrombospondins
Focal Adhesion Protein-Tyrosine Kinases
Focal Adhesions
Adhesion
Thrombospondin 1
Phosphatidylinositols
1-Phosphatidylinositol 4-Kinase
Phosphotransferases
Calreticulin
Phosphorylation
Chemical activation
Fibroblasts
LDL-Receptor Related Proteins
LDL Receptors
Endothelial cells
Cell Movement
Extracellular Matrix
Heparin

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Thrombospondin induces RhoA inactivation through FAK-dependent signaling to stimulate focal adhesion disassembly. / Orr, Anthony Wayne; Pallero, Manuel Antonio; Xiong, Wencheng; Murphy-Ullrich, Joanne E.

In: Journal of Biological Chemistry, Vol. 279, No. 47, 19.11.2004, p. 48983-48992.

Research output: Contribution to journalArticle

Orr, Anthony Wayne ; Pallero, Manuel Antonio ; Xiong, Wencheng ; Murphy-Ullrich, Joanne E. / Thrombospondin induces RhoA inactivation through FAK-dependent signaling to stimulate focal adhesion disassembly. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 47. pp. 48983-48992.
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