Rapid Remodeling of Invadosomes by Gi-coupled Receptors: DISSECTING THE ROLE OF Rho GTPases

Katarzyna M Kedziora, Daniela Leyton-Puig, Elisabetta Argenzio, Anja J Boumeester, Bram van Butselaar, Taofei Yin, Yi I Wu, Frank N van Leeuwen, Metello Innocenti, Kees Jalink, Wouter H Moolenaar

Research output: Contribution to journalArticlepeer-review

Abstract

Invadosomes are actin-rich membrane protrusions that degrade the extracellular matrix to drive tumor cell invasion. Key players in invadosome formation are c-Src and Rho family GTPases. Invadosomes can reassemble into circular rosette-like superstructures, but the underlying signaling mechanisms remain obscure. Here we show that Src-induced invadosomes in human melanoma cells (A375M and MDA-MB-435) undergo rapid remodeling into dynamic extracellular matrix-degrading rosettes by distinct G protein-coupled receptor agonists, notably lysophosphatidic acid (LPA; acting through the LPA1 receptor) and endothelin. Agonist-induced rosette formation is blocked by pertussis toxin, dependent on PI3K activity and accompanied by localized production of phosphatidylinositol 3,4,5-trisphosphate, whereas MAPK and Ca(2+) signaling are dispensable. Using FRET-based biosensors, we show that LPA and endothelin transiently activate Cdc42 through Gi, concurrent with a biphasic decrease in Rac activity and differential effects on RhoA. Cdc42 activity is essential for rosette formation, whereas G12/13-mediated RhoA-ROCK signaling suppresses the remodeling process. Our results reveal a Gi-mediated Cdc42 signaling axis by which G protein-coupled receptors trigger invadosome remodeling, the degree of which is dictated by the Cdc42-RhoA activity balance.

Original languageEnglish
Pages (from-to)4323-33
Number of pages11
JournalThe Journal of biological chemistry
Volume291
Issue number9
DOIs
Publication statusPublished - 26 Feb 2016
Externally publishedYes

Keywords

  • Biomarkers/metabolism
  • Cell Line, Tumor
  • Endothelins/metabolism
  • Extracellular Matrix/metabolism
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Hydrolysis
  • Luminescent Proteins/genetics
  • Lysophospholipids/metabolism
  • Melanoma/enzymology
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Neoplasm Proteins/agonists
  • Podosomes/enzymology
  • RNA Interference
  • Receptors, G-Protein-Coupled/antagonists & inhibitors
  • Receptors, Lysophosphatidic Acid/agonists
  • Recombinant Proteins/genetics
  • Time-Lapse Imaging
  • cdc42 GTP-Binding Protein/agonists
  • rac1 GTP-Binding Protein/agonists
  • rhoA GTP-Binding Protein/antagonists & inhibitors

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