title: The microtubule-associated protein EB1 maintains cell polarity through activation of protein kinase C.
abstract: The plus-ends of microtubules target the cell cortex to modulate actin protrusion dynamics and polarity, but little is known of the molecular mechanism that couples the interaction. EB1 protein associates with the plus-ends of microtubules, placing EB1 in an ideal spatial position to mediate microtubule-actin cross talk. The objective of the current study was to further understand intracellular signaling involved in EB1-dependent cell polarity and motility. B16F10 mouse melanoma cells were depleted of EB1 protein using short hair-pin RNA interference. Correlative live cell-immunofluorescence microscopy was performed to determine localization of WAVE2 and IQGAP1 to protruding versus retracting edges. EB1 knock down caused poor subcellular separation of WAVE2 and IQGAP1, and overall decreased localization. Activation of PKC corrected defects in WAVE2 and IQGAP1 localization, cell spreading and cell shape to levels observed in control cells, but did not correct defects in cell migration. Consistent with these findings, decreased PKC phosphorylation was observed in EB1 knock down cells. These findings support a model where EB1 protein links microtubules to actin protrusion and cell polarity through signaling pathways involving PKC.
authors: Department of Pharmaceutical Sciences, Southern Illinois University Edwardsville School of Pharmacy, Edwardsville, IL 62026-2000, USA. joschob@siue.edu
authors: Joseph M
authors: JM
authors: Schober
authors: Guim
authors: G
authors: Kwon
authors: Debbie
authors: D
authors: Jayne
authors: Jeanine M
authors: JM
authors: Cain
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002465:Cell Movement; D016764:Cell Polarity; D004789:Enzyme Activation; D055785:Gene Knockdown Techniques; D008546:Melanoma, Experimental / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D011493:Protein Kinase C / Q000096:biosynthesis*; D051300:Wiskott-Aldrich Syndrome Protein Family / Q000378:metabolism; D020703:ras GTPase-Activating Proteins / Q000378:metabolism
chemical_list: C405804:EB1 microtubule binding proteins; C101203:IQ motif containing GTPase activating protein 1; D008869:Microtubule-Associated Proteins; C492303:Wasf2 protein, mouse; D051300:Wiskott-Aldrich Syndrome Protein Family; D020703:ras GTPase-Activating Proteins; D011493:Protein Kinase C