TY - JOUR
T1 - Rapid Remodeling of Invadosomes by Gi-coupled Receptors
T2 - DISSECTING THE ROLE OF Rho GTPases
AU - Kedziora, Katarzyna M
AU - Leyton-Puig, Daniela
AU - Argenzio, Elisabetta
AU - Boumeester, Anja J
AU - van Butselaar, Bram
AU - Yin, Taofei
AU - Wu, Yi I
AU - van Leeuwen, Frank N
AU - Innocenti, Metello
AU - Jalink, Kees
AU - Moolenaar, Wouter H
N1 - © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2016/2/26
Y1 - 2016/2/26
N2 - 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.
AB - 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.
KW - Biomarkers/metabolism
KW - Cell Line, Tumor
KW - Endothelins/metabolism
KW - Extracellular Matrix/metabolism
KW - Fluorescence Resonance Energy Transfer
KW - Humans
KW - Hydrolysis
KW - Luminescent Proteins/genetics
KW - Lysophospholipids/metabolism
KW - Melanoma/enzymology
KW - Microscopy, Confocal
KW - Microscopy, Fluorescence
KW - Neoplasm Proteins/agonists
KW - Podosomes/enzymology
KW - RNA Interference
KW - Receptors, G-Protein-Coupled/antagonists & inhibitors
KW - Receptors, Lysophosphatidic Acid/agonists
KW - Recombinant Proteins/genetics
KW - Time-Lapse Imaging
KW - cdc42 GTP-Binding Protein/agonists
KW - rac1 GTP-Binding Protein/agonists
KW - rhoA GTP-Binding Protein/antagonists & inhibitors
UR - http://www.scopus.com/inward/record.url?scp=84966415524&partnerID=8YFLogxK
U2 - 10.1074/jbc.M115.695940
DO - 10.1074/jbc.M115.695940
M3 - Article
C2 - 26740622
SN - 0021-9258
VL - 291
SP - 4323
EP - 4333
JO - The Journal of biological chemistry
JF - The Journal of biological chemistry
IS - 9
ER -