The anchoring protein RACK1 links protein kinase Cepsilon to integrin beta chains. Requirements for adhesion and motility.
Integrin affinity is modulated by intracellular signaling cascades, in a process known as "inside-out" signaling, leading to changes in cell adhesion and motility. Protein kinase C (PKC) plays a critical role in integrin-mediated events; however, the mechanism that links PKC to integrins remains unclear. Here, we report that PKCepsilon positively regulates integrin-dependent adhesion, spreading, and motility of human glioma cells. PKCepsilon activation was associated with increased focal adhesion and lamellipodia formation as well as clustering of select integrins, and it is required for phorbol 12-myristate 13-acetate-induced adhesion and motility. We provide novel evidence that the scaffolding protein RACK1 mediates the interaction between integrin beta chain and activated PKCepsilon. Both depletion of RACK1 by antisense strategy and overexpression of a truncated form of RACK1 which lacks the integrin binding region resulted in decreased PKCepsilon-induced adhesion and migration, suggesting that RACK1 links PKCepsilon to integrin beta chains. Altogether, these results provide a novel mechanistic link between PKC activation and integrin-mediated adhesion and motility.
Besson
Arnaud
A
Department of Oncology, University of Calgary, Calgary, Alberta T2N 4N1, Canada.
Wilson
Tammy L
TL
Yong
V Wee
VW
D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D002448:Cell Adhesion; D002465:Cell Movement; D018076:DNA, Complementary / Q000378:metabolism; D004789:Enzyme Activation; D019204:GTP-Binding Proteins; D006801:Humans; D016023:Integrins / Q000737:chemistry / Q000378:metabolism*; D007527:Isoenzymes / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D009363:Neoplasm Proteins / Q000737:chemistry* / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D011493:Protein Kinase C / Q000737:chemistry* / Q000378:metabolism; D051571:Protein Kinase C-alpha; D051744:Protein Kinase C-epsilon; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; D013347:Subcellular Fractions; D013997:Time Factors; D014162:Transfection; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D016023:Integrins; D007527:Isoenzymes; D009363:Neoplasm Proteins; C419368:RACK1 protein, human; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; C497256:Prkce protein, mouse; C496628:PRKCA protein, human; C497255:PRKCE protein, human; C496630:Prkca protein, mouse; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D051744:Protein Kinase C-epsilon; D019204:GTP-Binding Proteins