title: Epidermal growth factor-induced cellular invasion requires sphingosine-1-phosphate/sphingosine-1-phosphate 2 receptor-mediated ezrin activation.
abstract: Ezrin, radixin, and moesin (ERM) proteins link cortical actin to the plasma membrane and coordinate cellular events that require cytoskeletal rearrangement, including cell division, migration, and invasion. While ERM proteins are involved in many important cellular events, the mechanisms regulating their function are not completely understood. Our laboratory previously identified reciprocal roles for the sphingolipids ceramide and sphingosine-1-phosphate (S1P) in the regulation of ERM proteins. We recently showed that ceramide-induced activation of PP1α leads to dephosphorylation and inactivation of ERM proteins, while S1P results in phosphorylation and activation of ERM proteins. Following these findings, we aimed to examine known inducers of the SK/S1P pathway and evaluate their ability to regulate ERM proteins. We examined EGF, a known inducer of the SK/S1P pathway, for its ability to regulate the ERM family of proteins. We found that EGF induces ERM c-terminal threonine phosphorylation via activation of the SK/S1P pathway, as this was prevented by siRNA knockdown or pharmacological inhibition of SK. Using pharmacological, as well as genetic, knockdown approaches, we determined that EGF induces ERM phosphorylation via activation of S1PR2. In addition, EGF led to cell polarization in the form of lamellipodia, and this occurred through a mechanism involving S1PR2-mediated phosphorylation of ezrin T567. EGF-induced cellular invasion was also found to be dependent on S1PR2-induced T567 ezrin phosphorylation, such that S1PR2 antagonist, JTE-013, and expression of a dominant-negative ezrin mutant prevented cellular invasion toward EGF. In this work, a novel mechanism of EGF-stimulated invasion is unveiled, whereby S1P-mediated activation of S1PR2 and phosphorylation of ezrin T567 is required.
authors: Department of Molecular and Cellular Biology and Pathobiology, Medical University of South Carolina, Charleston, South Carolina, USA.
authors: K Alexa
authors: KA
authors: Orr Gandy
authors: Mohamad
authors: M
authors: Adada
authors: Daniel
authors: D
authors: Canals
authors: Brittany
authors: B
authors: Carroll
authors: Patrick
authors: P
authors: Roddy
authors: Yusuf A
authors: YA
authors: Hannun
authors: Lina M
authors: LM
authors: Obeid
mesh_terms: D002465:Cell Movement / Q000187:drug effects / Q000235:genetics; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D008246:Lysophospholipids / Q000378:metabolism*; D018613:Microscopy, Confocal; D009154:Mutation; D010766:Phosphorylation / Q000187:drug effects; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism*; D011720:Pyrazoles / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D034622:RNA Interference; D049349:Receptors, Lysosphingolipid / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000378:metabolism; D000081025:Sphingosine-1-Phosphate Receptors; D000099157:Sphingosine Kinase; D000099149:Ezrin
chemical_list: D003598:Cytoskeletal Proteins; D004815:Epidermal Growth Factor; D008246:Lysophospholipids; D017853:Phosphotransferases (Alcohol Group Acceptor); D011720:Pyrazoles; D011725:Pyridines; D049349:Receptors, Lysosphingolipid; D013110:Sphingosine; D000081025:Sphingosine-1-Phosphate Receptors; D000099157:Sphingosine Kinase; D000099149:Ezrin; C471998:JTE 013; C477522:S1PR2 protein, human; C060506:sphingosine 1-phosphate
keywords: actin cytoskeleton; moesin; polarization; radixin
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