Rac1-dependent phosphorylation and focal adhesion recruitment of myosin IIA regulates migration and mechanosensing

Curr Biol. 2015 Jan 19;25(2):175-186. doi: 10.1016/j.cub.2014.11.043. Epub 2014 Dec 24.

Abstract

Background: Cell migration requires coordinated formation of focal adhesions (FAs) and assembly and contraction of the actin cytoskeleton. Nonmuscle myosin II (MII) is a critical mediator of contractility and FA dynamics in cell migration. Signaling downstream of the small GTPase Rac1 also regulates FA and actin dynamics, but its role in regulation of MII during migration is less clear.

Results: We found that Rac1 promotes association of MIIA with FA. Live-cell imaging showed that, whereas most MIIA at the leading edge assembled into dorsal contractile arcs, a substantial subset assembled in or was captured within maturing FA, and this behavior was promoted by active Rac1. Protein kinase C (PKC) activation was necessary and sufficient for integrin- and Rac1-dependent phosphorylation of MIIA heavy chain (HC) on serine1916 (S1916) and recruitment to FA. S1916 phosphorylation of MIIA HC and localization in FA was enhanced during cell spreading and ECM stiffness mechanosensing, suggesting upregulation of this pathway during physiological Rac1 activation. Phosphomimic and nonphosphorylatable MIIA HC mutants demonstrated that S1916 phosphorylation was necessary and sufficient for the capture and assembly of MIIA minifilaments in FA. S1916 phosphorylation was also sufficient to promote the rapid assembly of FAs to enhance cell migration and for the modulation of traction force, spreading, and migration by ECM stiffness.

Conclusions: Our study reveals for the first time that Rac1 and integrin activation regulates MIIA HC phosphorylation through a PKC-dependent mechanism that promotes MIIA association with FAs and acts as a critical modulator of cell migration and mechanosensing.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, N.I.H., Intramural

MeSH terms

  • Cell Line
  • Cell Movement*
  • Focal Adhesions / metabolism*
  • Humans
  • Mechanotransduction, Cellular / physiology
  • Molecular Motor Proteins / genetics*
  • Molecular Motor Proteins / metabolism
  • Myosin Heavy Chains / genetics*
  • Myosin Heavy Chains / metabolism
  • Phosphorylation
  • Signal Transduction*
  • rac1 GTP-Binding Protein / genetics*
  • rac1 GTP-Binding Protein / metabolism

Substances

  • MYH9 protein, human
  • Molecular Motor Proteins
  • RAC1 protein, human
  • Myosin Heavy Chains
  • rac1 GTP-Binding Protein