Abstract
Dvl is a key protein that transmits the Wnt signal to the canonical beta-catenin pathway and the noncanonical planar cell polarity (PCP) pathway. We studied the roles of Rho-associated kinase (Rho-kinase), which is activated by Dvl in the PCP pathway of mammalian cells. The expression of Dvl-1, Wnt-1, or Wnt-3a activated Rho-kinase in COS cells, and this activation was inhibited by the Rho-binding domain of Rho-kinase. The expression of Dvl-1 in PC12 cells activated Rho and inhibited nerve growth factor (NGF)-induced neurite outgrowth. This inhibition was reversed by a Rho-kinase inhibitor but not by a c-Jun N-terminal kinase inhibitor. Dvl-1 also inhibited serum starvation-dependent neurite outgrowth of N1E-115 cells, and expression of the Rho-binding domain of Rho-kinase reversed this inhibitory activity of Dvl-1. Dvl-1 mutants that did not activate Rho-kinase did not inhibit the neurite outgrowth of N1E-115 cells. Furthermore, the purified Wnt-3a protein activated Rho-kinase and inhibited the NGF-dependent neurite outgrowth of PC12 cells. Wnt-3a-dependent neurite retraction was also prevented by a Rho-kinase inhibitor and a Dvl-1 mutant that suppresses Wnt-3a-dependent activation of Rho-kinase. These results suggest that Wnt-3a and Dvl regulate neurite formation through Rho-kinase and that PC12 and N1E-115 cells are useful for analyzing the PCP pathway.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing
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Animals
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COS Cells
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Cattle
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Cell Line
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Cell Polarity
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Culture Media, Serum-Free
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Dishevelled Proteins
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Enzyme Activation
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Enzyme Inhibitors / pharmacology
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Glycogen Synthase Kinase 3 / antagonists & inhibitors
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Humans
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Intracellular Signaling Peptides and Proteins
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JNK Mitogen-Activated Protein Kinases
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Mice
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Mitogen-Activated Protein Kinases / antagonists & inhibitors
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Nerve Growth Factor / pharmacology
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Neurites / drug effects
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Neurites / physiology*
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Neurons / metabolism
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Neurons / ultrastructure
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PC12 Cells
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Phosphoproteins / genetics
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Phosphoproteins / metabolism*
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Protein Serine-Threonine Kinases / genetics
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Protein Serine-Threonine Kinases / metabolism*
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Proteins / genetics
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Proteins / metabolism*
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Proteins / pharmacology
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Rats
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Signal Transduction
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Wnt Proteins
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Wnt3 Protein
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Wnt3A Protein
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rho-Associated Kinases
Substances
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Adaptor Proteins, Signal Transducing
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Culture Media, Serum-Free
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DVL1 protein, human
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Dishevelled Proteins
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Dvl1 protein, mouse
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Dvl1 protein, rat
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Enzyme Inhibitors
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Intracellular Signaling Peptides and Proteins
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Phosphoproteins
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Proteins
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Recombinant Fusion Proteins
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WNT3A protein, human
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Wnt Proteins
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Wnt3 Protein
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Wnt3A Protein
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Wnt3a protein, mouse
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Nerve Growth Factor
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Protein Serine-Threonine Kinases
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rho-Associated Kinases
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JNK Mitogen-Activated Protein Kinases
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Mitogen-Activated Protein Kinases
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Glycogen Synthase Kinase 3