title: Rapid Remodeling of Invadosomes by Gi-coupled Receptors: DISSECTING THE ROLE OF Rho GTPases.
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.
authors.affiliation: From the Division of Cell Biology and.
authors.forename: Katarzyna M
authors.initials: KM
authors.lastname: Kedziora
authors.affiliation: From the Division of Cell Biology and.
authors.forename: Daniela
authors.initials: D
authors.lastname: Leyton-Puig
authors.affiliation: From the Division of Cell Biology and.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Argenzio
authors.affiliation: From the Division of Cell Biology and.
authors.forename: Anja J
authors.initials: AJ
authors.lastname: Boumeester
authors.affiliation: From the Division of Cell Biology and.
authors.forename: Bram
authors.initials: B
authors.lastname: van Butselaar
authors.affiliation: the Center for Cell Analysis and Modeling, University of Connecticut Health Center, Farmington, Connecticut 06030, and.
authors.forename: Taofei
authors.initials: T
authors.lastname: Yin
authors.affiliation: the Center for Cell Analysis and Modeling, University of Connecticut Health Center, Farmington, Connecticut 06030, and.
authors.forename: Yi I
authors.initials: YI
authors.lastname: Wu
authors.affiliation: the Department of Cell Biology, Radboud University Medical Centre, 6525 GA Nijmegen, The Netherlands.
authors.forename: Frank N
authors.initials: FN
authors.lastname: van Leeuwen
authors.affiliation: Division of Molecular Genetics, Netherlands Cancer Institute, Amsterdam 1066 CX, The Netherlands.
authors.forename: Metello
authors.initials: M
authors.lastname: Innocenti
authors.affiliation: From the Division of Cell Biology and k.jalink@nki.nl.
authors.forename: Kees
authors.initials: K
authors.lastname: Jalink
authors.affiliation: From the Division of Cell Biology and.
authors.forename: Wouter H
authors.initials: WH
authors.lastname: Moolenaar
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D045744:Cell Line, Tumor; D016232:Endothelins / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism / Q000473:pathology; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D006868:Hydrolysis; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008246:Lysophospholipids / Q000378:metabolism*; D008545:Melanoma / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009363:Neoplasm Proteins / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000069261:Podosomes / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D034622:RNA Interference; D043562:Receptors, G-Protein-Coupled / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D049368:Receptors, Lysophosphatidic Acid / Q000819:agonists* / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D059008:Time-Lapse Imaging; D020764:cdc42 GTP-Binding Protein / Q000819:agonists* / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D020830:rac1 GTP-Binding Protein / Q000819:agonists / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: D015415:Biomarkers; D016232:Endothelins; C489011:GPR26 protein, human; D008164:Luminescent Proteins; D008246:Lysophospholipids; D009363:Neoplasm Proteins; C107953:RAC1 protein, human; D043562:Receptors, G-Protein-Coupled; D049368:Receptors, Lysophosphatidic Acid; D011994:Recombinant Proteins; C066737:RHOA protein, human; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein; D020742:rhoA GTP-Binding Protein; C032881:lysophosphatidic acid
keywords: CDC42; G protein-coupled receptor (GPCR); Rac (Rac GTPase); Rho (Rho GTPase); biosensor; calcium intracellular release; fluorescence resonance energy transfer (FRET); imaging; invadopodia; phosphatidylinositide 3-kinase (PI 3-kinase)