Integrin alpha3beta1-dependent activation of FAK/Src regulates Rac1-mediated keratinocyte polarization on laminin-5.
Upon epidermal wounding, keratinocytes at the wound edge become activated, deposit newly synthesized laminin-5 into the extracellular matrix, and migrate into the wound bed. The interaction between integrin alpha3beta1 and laminin-5 is essential for establishment of a stable, leading lamellipodium and persistent keratinocyte migration. We previously showed that integrin alpha3beta1 activates the Rho family GTPase Rac1 and regulates Rac1-dependent formation of polarized, leading lamellipodia in migrating keratinocytes. In the present study, we explored the role of focal adhesion kinase (FAK) and src signaling in this process. We show that overexpression of the FAK inhibitor FAK-related non-kinase or of the FAK(Y397F) auto-phosphorylation mutant, induced abnormal, non-polarized spreading of keratinocytes on laminin-5. Integrin alpha3beta1 was required for full FAK auto-phosphorylation at Y397, and subsequent src kinase-dependent phosphorylation of FAK at residues Y861 and Y925, sites responsible for promoting signal transduction downstream of FAK, indicating that alpha3beta1 regulates the coordination of FAK/src signal transduction. Inhibiting either src kinase activity or FAK signaling interfered with alpha3beta1-mediated Rac1 activation and polarized cell spreading. These findings reveal a novel pathway in migratory keratinocytes wherein alpha3beta1-laminin-5 interactions regulate src kinase signaling through FAK, promoting Rac1 activation and polarized lamellipodium extension.
Choma
David P
DP
Center for Cell Biology and Cancer Research, Albany Medical College, Albany, New York 12208-3479, USA.
Milano
Vincenzo
V
Pumiglia
Kevin M
KM
DiPersio
C Michael
CM
D000818:Animals; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000502:physiology*; D016764:Cell Polarity; D002478:Cells, Cultured; D048049:Extracellular Signal-Regulated MAP Kinases / Q000502:physiology; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000502:physiology*; D039201:Integrin alpha3beta1 / Q000502:physiology*; D015603:Keratinocytes / Q000502:physiology*; D051379:Mice; D010766:Phosphorylation; D011743:Pyrimidines / Q000494:pharmacology; D015398:Signal Transduction; D020830:rac1 GTP-Binding Protein / Q000502:physiology*; D019061:src-Family Kinases / Q000502:physiology*; D000097794:Kalinin
D015815:Cell Adhesion Molecules; D048049:Extracellular Signal-Regulated MAP Kinases; D051416:Focal Adhesion Protein-Tyrosine Kinases; D039201:Integrin alpha3beta1; D011743:Pyrimidines; D020830:rac1 GTP-Binding Protein; D019061:src-Family Kinases; D000097794:Kalinin; C412373:AG 1879