FARP2 triggers signals for Sema3A-mediated axonal repulsion

Sema3A, a prototypical semaphorin, acts as a chemorepellent or a chemoattractant for axons by activating a receptor complex comprising neuropilin-1 as the ligand-binding subunit and plexin-A1 as the signal-transducing subunit. How the signals downstream of plexin-A1 are triggered upon Sema3A stimulation, however, is unknown. Here we show that, in the presence of neuropilin-1, the FERM domain–containing guanine nucleotide exchange factor (GEF) FARP2 associates directly with plexin-A1. Sema3A binding to neuropilin-1 induces the dissociation of FARP2 from plexin-A1, resulting in activation of FARP2's Rac GEF activity, Rnd1 recruitment to plexin-A1, and downregulation of R-Ras. Simultaneously, the FERM domain of FARP2 sequesters phosphatidylinositol phosphate kinase type I isoform PIPKIγ661 from talin, thereby inhibiting its kinase activity. These activities are required for Sema3A-mediated repulsion of outgrowing axons and suppression of neuronal adhesion. We therefore conclude that FARP2 is a key molecule involved in the response of neuronal growth cones to class-3 semaphorins.

[1]  Fumio Nakamura,et al.  Collapsin-induced growth cone collapse mediated by an intracellular protein related to UNC-33 , 1995, Nature.

[2]  Tianyi Mao,et al.  MICALs, a Family of Conserved Flavoprotein Oxidoreductases, Function in Plexin-Mediated Axonal Repulsion , 2002, Cell.

[3]  M. Tessier-Lavigne,et al.  Class 3 semaphorins control vascular morphogenesis by inhibiting integrin function , 2003, Nature.

[4]  Toshihide Yamashita,et al.  A Novel FERM Domain Including Guanine Nucleotide Exchange Factor Is Involved in Rac Signaling and Regulates Neurite Remodeling , 2002, The Journal of Neuroscience.

[5]  Hironori Katoh,et al.  Direct Interaction of Rnd1 with Plexin-B1 Regulates PDZ-RhoGEF-mediated Rho Activation by Plexin-B1 and Induces Cell Contraction in COS-7 Cells* , 2003, Journal of Biological Chemistry.

[6]  B. Eickholt,et al.  Sema3A-induced growth-cone collapse is mediated by Rac1 amino acids 17–32 , 1999, Current Biology.

[7]  Frits Michiels,et al.  Matrix-dependent Tiam1/Rac Signaling in Epithelial Cells Promotes Either Cell–Cell Adhesion or Cell Migration and Is Regulated by Phosphatidylinositol 3-Kinase , 1998, The Journal of cell biology.

[8]  G. Gallo,et al.  Axon guidance: GTPases help axons reach their targets , 1998, Current Biology.

[9]  C. Goodman,et al.  The Molecular Biology of Axon Guidance , 1996, Science.

[10]  M. Hori,et al.  Dual roles of Sema6D in cardiac morphogenesis through region-specific association of its receptor, Plexin-A1, with off-track and vascular endothelial growth factor receptor type 2. , 2004, Genes & development.

[11]  P. Forscher,et al.  Substrate-cytoskeletal coupling as a mechanism for the regulation of growth cone motility and guidance. , 2000, Journal of neurobiology.

[12]  Zhigang He,et al.  Semaphorin–Neuropilin Interactions Underlying Sympathetic Axon Responses to Class III Semaphorins , 1998, Neuron.

[13]  A. Kolodkin,et al.  Semaphorin junction: making tracks toward neural connectivity , 2003, Current Opinion in Neurobiology.

[14]  I. Hovatta,et al.  The semaphorin 3A receptor may directly regulate the activity of small GTPases , 2000, FEBS letters.

[15]  P. Grabham,et al.  Nerve growth factor stimulates coupling of beta1 integrin to distinct transport mechanisms in the filopodia of growth cones. , 2000, Journal of cell science.

[16]  Kristiina Vuori,et al.  Integrin Activation by R-ras , 1996, Cell.

[17]  Y. Zheng,et al.  Guanine nucleotide exchange catalyzed by dbl oncogene product. , 1995, Methods in enzymology.

[18]  S. Offermanns,et al.  Plexin-B1 Directly Interacts with PDZ-RhoGEF/LARG to Regulate RhoA and Growth Cone Morphology , 2002, Neuron.

[19]  D. Webb,et al.  Talin: an emerging focal point of adhesion dynamics. , 2004, Current opinion in cell biology.

[20]  Pietro De Camilli,et al.  Recruitment and regulation of phosphatidylinositol phosphate kinase type 1γ by the FERM domain of talin , 2002, Nature.

[21]  P. Keely,et al.  R-Ras Signals through Specific Integrin α Cytoplasmic Domains to Promote Migration and Invasion of Breast Epithelial Cells , 1999, The Journal of cell biology.

[22]  H. Vikis,et al.  The plexin-B1/Rac interaction inhibits PAK activation and enhances Sema4D ligand binding. , 2002, Genes & development.

[23]  M. vanTriest,et al.  Measurement of GTP-bound Ras-like GTPases by activation-specific probes. , 2001 .

[24]  M. Greenberg,et al.  Orchestral Maneuvers in the Axon Trio and the Control of Axon Guidance , 2000, Cell.

[25]  L. Lim,et al.  Rho family GTPases and neuronal growth cone remodelling: relationship between increased complexity induced by Cdc42Hs, Rac1, and acetylcholine and collapse induced by RhoA and lysophosphatidic acid , 1997, Molecular and cellular biology.

[26]  J. Bos,et al.  Measurement of GTP-bound Ras-like GTPases by activation-specific probes. , 2001, Methods in enzymology.

[27]  P. Grabham,et al.  Nerve Growth Factor Stimulates the Accumulation of β1 Integrin at the Tips of Filopodia in the Growth Cones of Sympathetic Neurons , 1997, The Journal of Neuroscience.

[28]  N. Ahn,et al.  The semaphorin receptor plexin-B1 signals through a direct interaction with the Rho-specific nucleotide exchange factor, LARG , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[29]  K. Ling,et al.  Type I gamma phosphatidylinositol phosphate kinase targets and regulates focal adhesions. , 2002, Nature.

[30]  K. Ling,et al.  Type Iγ phosphatidylinositol phosphate kinase targets and regulates focal adhesions , 2002, Nature.

[31]  E. Maestrini,et al.  A family of transmembrane proteins with homology to the MET-hepatocyte growth factor receptor. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[32]  J. Gutkind,et al.  Plexin B Regulates Rho through the Guanine Nucleotide Exchange Factors Leukemia-associated Rho GEF (LARG) and PDZ-RhoGEF* , 2002, The Journal of Biological Chemistry.

[33]  H. Katoh,et al.  The Semaphorin 4D Receptor Plexin-B1 Is a GTPase Activating Protein for R-Ras , 2004, Science.

[34]  H. Cline,et al.  Regulation of Rho GTPases by Crosstalk and Neuronal Activity In Vivo , 2002, Neuron.

[35]  L. Luo.  RHO GTPASES in neuronal morphogenesis , 2000, Nature Reviews Neuroscience.

[36]  A. Hall,et al.  The Activity of the Plexin-A1 Receptor Is Regulated by Rac* , 2004, Journal of Biological Chemistry.

[37]  P. Mangeat,et al.  ERM proteins in cell adhesion and membrane dynamics. , 1999, Trends in cell biology.

[38]  C. Goodman,et al.  Bi-directional signaling by Semaphorin 1a during central synapse formation in Drosophila , 2002, Nature Neuroscience.

[39]  Hironori Katoh,et al.  Molecular Dissection of the Semaphorin 4D Receptor Plexin-B1-Stimulated R-Ras GTPase-Activating Protein Activity and Neurite Remodeling in Hippocampal Neurons , 2004, The Journal of Neuroscience.

[40]  S. Strittmatter,et al.  Rac1 Mediates Collapsin-1-Induced Growth Cone Collapse , 1997, The Journal of Neuroscience.

[41]  M. Hori,et al.  Guidance of myocardial patterning in cardiac development by Sema6D reverse signalling , 2004, Nature Cell Biology.

[42]  S. Zanata,et al.  Antagonistic Effects of Rnd1 and RhoD GTPases Regulate Receptor Activity in Semaphorin 3A-Induced Cytoskeletal Collapse , 2002, The Journal of Neuroscience.

[43]  C. Wilcox,et al.  Myelin and Collapsin-1 Induce Motor Neuron Growth Cone Collapse through Different Pathways: Inhibition of Collapse by Opposing Mutants of Rac1 , 1999, The Journal of Neuroscience.

[44]  C. Isacke,et al.  Identification and functional analysis of the ezrin-binding site in the hyaluronan receptor, CD44 , 1998, Current Biology.