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Area of Science:

  • Cell Biology
  • Mechanobiology
  • Biophysics

Background:

  • Cells interact with their environment via membrane protrusion-retraction cycles.
  • Rac1 and RhoA GTP-binding activities coordinate these cycles, but their lifetime control is unknown.

Purpose of the Study:

  • Investigate the relationship between Rac1/RhoA GTP-binding levels and protrusion/retraction events.
  • Examine the impact on cell-extracellular matrix (ECM) traction forces at relevant ECM stiffness.

Main Methods:

  • Measured Rac1 and RhoA GTP-binding levels.
  • Analyzed protrusion and retraction events.
  • Assessed cell-ECM traction forces.
  • Utilized optogenetics for local Rac1-GTP perturbations.

Main Results:

  • High RhoA-GTP preceded retractions; Rac1-GTP elevation preceded protrusions.
  • Rac1-GTP levels correlated with maximal edge velocity during protrusions and retractions.
  • Altered Rac1-GTP preceded changes in traction force.
  • Optogenetic Rac1-GTP manipulation showed Rac1 restricts protrusions/retractions and promotes traction.

Conclusions:

  • Rac1 plays a key role in limiting the duration and size of membrane protrusions and retractions.
  • Rac1 influences cell-ECM traction forces, contributing to the regulation of cell movement.