Adhesion-dependent regulation of p190RhoGAP in the developing brain by the Abl-related gene tyrosine kinase.
Abl family kinases, which include the mammalian Abl and Arg (Abl-related gene) kinases, regulate neuronal morphogenesis in developing metazoa (for review, see [1]). Activation of Abl kinase activity directs changes in actin-dependent processes such as membrane ruffling, filopodial protrusion, and cell motility. However, the mechanisms by which increased Abl or Arg kinase activity promote cytoskeletal rearrangements are unclear. We provide evidence that the Rho inhibitor p190RhoGAP (GTPase-activating protein) is an Arg substrate in the postnatal mouse brain. We show that p190RhoGAP has reduced phosphotyrosine content in postnatal arg(-/-) mouse brain extracts relative to wild-type extracts. In addition, the adhesion-dependent stimulation of p190RhoGAP phosphorylation observed in wild-type cells is not observed in arg(-/-) fibroblasts and neurons. Arg can phosphorylate p190RhoGAP in vitro and in vivo on tyrosine (Y) 1105. We find that Arg can stimulate p190RhoGAP to inhibit Rho and that Arg-mediated phosphorylation is required for this stimulation. Phosphorylation by Arg also promotes p190RhoGAP's association with p120RasGAP and stimulates p190RhoGAP's ability to induce neuritogenesis in neuroblastoma cells. Our results demonstrate that p190RhoGAP is an Arg substrate in the developing brain and suggest that Arg mediates the adhesion-dependent regulation of neuronal morphogenesis in the postnatal brain by phosphorylating p190RhoGAP.
Hernández
Samuel E
SE
Interdepartmental Neuroscience Program, Yale University, New Haven, CT 06520, USA.
Settleman
Jeffrey
J
Koleske
Anthony J
AJ
D016475:3T3 Cells; D000818:Animals; D001921:Brain / Q000196:embryology* / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism*; D004268:DNA-Binding Proteins; D020690:GTPase-Activating Proteins; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D015151:Immunoblotting; D051379:Mice; D009024:Morphogenesis; D016501:Neurites / Q000502:physiology; D009474:Neurons / Q000502:physiology*; D009687:Nuclear Proteins / Q000378:metabolism*; D010766:Phosphorylation; D010957:Plasmids; D011233:Precipitin Tests; D011505:Protein-Tyrosine Kinases / Q000378:metabolism* / Q000502:physiology; D012097:Repressor Proteins; D014162:Transfection; D014407:Tumor Cells, Cultured
C513041:Arhgap35 protein, mouse; C513042:Arhgap5 protein, mouse; D004268:DNA-Binding Proteins; D020690:GTPase-Activating Proteins; D020662:Guanine Nucleotide Exchange Factors; D009687:Nuclear Proteins; D012097:Repressor Proteins; C426236:ARG tyrosine kinase; D011505:Protein-Tyrosine Kinases