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Requirement for Rho in integrin signalling

S T Barry1, H M Flinn, M J Humphries

  • 1Department of Biochemistry, University of Leicester, UK.

Cell Adhesion and Communication
|March 1, 1997
PubMed
Summary
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The small GTP-binding protein Rho is crucial for forming focal adhesions and actin stress fibers in Swiss 3T3 cells. Rho acts downstream of integrin signaling, mediating cell adhesion and cytoskeletal organization.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Serum-free culture of Swiss 3T3 cells causes loss of focal adhesions and actin stress fibers.
  • The small GTP-binding protein Rho is essential for growth factor-induced stress fiber and focal adhesion formation.

Purpose of the Study:

  • To investigate the role of Rho in integrin-mediated focal adhesion and stress fiber assembly.
  • To determine if Rho signaling is downstream of integrin activation.

Main Methods:

  • Swiss 3T3 cells cultured in serum-free medium.
  • Stimulation with lysophosphatidic acid (LPA) and extracellular matrix proteins.
  • Inhibition of Rho using C3 transferase via scrape-loading and microinjection.
  • Analysis of focal adhesions, actin stress fibers, and protein tyrosine phosphorylation.

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Main Results:

  • Rho is required for LPA-induced stress fibers, focal adhesions, and tyrosine phosphorylation of focal adhesion proteins.
  • Plating on extracellular matrix proteins induced tyrosine phosphorylation, stress fibers, and focal adhesions, inhibited by C3 transferase.
  • GRGDS peptide triggered focal adhesion and stress fiber formation, blocked by C3 transferase, indicating Rho-dependent integrin signaling.

Conclusions:

  • Rho GTPase is a key mediator in the signaling pathway downstream of integrin activation.
  • Integrin engagement with extracellular matrix proteins stimulates Rho-dependent cytoskeletal organization and focal adhesion assembly.