RPEL-family rhoGAPs link Rac/Cdc42 GTP loading to G-actin availability

Nat Cell Biol. 2019 Jul;21(7):845-855. doi: 10.1038/s41556-019-0337-y. Epub 2019 Jun 17.

Abstract

RPEL proteins, which contain the G-actin-binding RPEL motif, coordinate cytoskeletal processes with actin dynamics. We show that the ArhGAP12- and ArhGAP32-family GTPase-activating proteins (GAPs) are RPEL proteins. We determine the structure of the ArhGAP12/G-actin complex, and show that G-actin contacts the RPEL motif and GAP domain sequences. G-actin inhibits ArhGAP12 GAP activity, and this requires the G-actin contacts identified in the structure. In B16 melanoma cells, ArhGAP12 suppresses basal Rac and Cdc42 activity, F-actin assembly, invadopodia formation and experimental metastasis. In this setting, ArhGAP12 mutants defective for G-actin binding exhibit more effective downregulation of Rac GTP loading following HGF stimulation and enhanced inhibition of Rac-dependent processes, including invadopodia formation. Potentiation or disruption of the G-actin/ArhGAP12 interaction, by treatment with the actin-binding drugs latrunculin B or cytochalasin D, has corresponding effects on Rac GTP loading. The interaction of G-actin with RPEL-family rhoGAPs thus provides a negative feedback loop that couples Rac activity to actin dynamics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / drug effects
  • Actins / metabolism*
  • Animals
  • Bridged Bicyclo Compounds, Heterocyclic / pharmacology
  • Cytochalasin D / pharmacology
  • GTPase-Activating Proteins / drug effects
  • GTPase-Activating Proteins / genetics
  • GTPase-Activating Proteins / metabolism*
  • Guanosine Triphosphate / metabolism
  • Humans
  • Mice
  • Protein Binding / drug effects
  • Protein Binding / genetics
  • Rabbits
  • Thiazolidines / pharmacology
  • cdc42 GTP-Binding Protein / drug effects
  • cdc42 GTP-Binding Protein / metabolism*
  • rac GTP-Binding Proteins / metabolism

Substances

  • ARHGAP12 protein, human
  • Actins
  • Bridged Bicyclo Compounds, Heterocyclic
  • GTPase-Activating Proteins
  • Thiazolidines
  • rho GTPase-activating protein
  • Cytochalasin D
  • Guanosine Triphosphate
  • cdc42 GTP-Binding Protein
  • rac GTP-Binding Proteins
  • latrunculin B