title: Phosphorylation by Rho kinase regulates CRMP-2 activity in growth cones.
abstract: Collapsin response mediator protein 2 (CRMP-2) enhances the advance of growth cones by regulating microtubule assembly and Numb-mediated endocytosis. We previously showed that Rho kinase phosphorylates CRMP-2 during growth cone collapse; however, the roles of phosphorylated CRMP-2 in growth cone collapse remain to be clarified. Here, we report that CRMP-2 phosphorylation by Rho kinase cancels the binding activity to the tubulin dimer, microtubules, or Numb. CRMP-2 binds to actin, but its binding is not affected by phosphorylation. Electron microscopy revealed that CRMP-2 localizes on microtubules, clathrin-coated pits, and actin filaments in dorsal root ganglion neuron growth cones, while phosphorylated CRMP-2 localizes only on actin filaments. The phosphomimic mutant of CRMP-2 has a weakened ability to enhance neurite elongation. Furthermore, ephrin-A5 induces phosphorylation of CRMP-2 via Rho kinase during growth cone collapse. Taken together, these results suggest that Rho kinase phosphorylates CRMP-2, and inactivates the ability of CRMP-2 to promote microtubule assembly and Numb-mediated endocytosis, during growth cone collapse.
authors.affiliation: Department of Cell Pharmacology, Graduate School of Medicine, Nagoya University, 65 Tsurumai, Showa, Nagoya, Aichi 466-8550, Japan.
authors.forename: Nariko
authors.initials: N
authors.lastname: Arimura
authors.forename: Céline
authors.initials: C
authors.lastname: Ménager
authors.forename: Yoji
authors.initials: Y
authors.lastname: Kawano
authors.forename: Takeshi
authors.initials: T
authors.lastname: Yoshimura
authors.forename: Saeko
authors.initials: S
authors.lastname: Kawabata
authors.forename: Atsushi
authors.initials: A
authors.lastname: Hattori
authors.forename: Yuko
authors.initials: Y
authors.lastname: Fukata
authors.forename: Mutsuki
authors.initials: M
authors.lastname: Amano
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Goshima
authors.forename: Masaki
authors.initials: M
authors.lastname: Inagaki
authors.forename: Nobuhiro
authors.initials: N
authors.lastname: Morone
authors.forename: Jiro
authors.initials: J
authors.lastname: Usukura
authors.forename: Kozo
authors.initials: K
authors.lastname: Kaibuchi
mesh_terms: D000818:Animals; D001369:Axons / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation; D002642:Chick Embryo; D002522:Chlorocebus aethiops; D018076:DNA, Complementary / Q000378:metabolism; D019281:Dimerization; D029721:Drosophila Proteins / Q000378:metabolism*; D004705:Endocytosis; D036386:Ephrin-A5 / Q000378:metabolism; D005727:Ganglia, Spinal / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D007150:Immunohistochemistry; D036341:Intercellular Signaling Peptides and Proteins; D047908:Intracellular Signaling Peptides and Proteins; D007605:Juvenile Hormones / Q000378:metabolism*; D051379:Mice; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism; D008954:Models, Biological; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D020349:Surface Plasmon Resonance; D013997:Time Factors; D014162:Transfection; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism; D014709:Vero Cells; D054460:rho-Associated Kinases
chemical_list: D018076:DNA, Complementary; D029721:Drosophila Proteins; D036386:Ephrin-A5; D036341:Intercellular Signaling Peptides and Proteins; D047908:Intracellular Signaling Peptides and Proteins; D007605:Juvenile Hormones; D009419:Nerve Tissue Proteins; D014404:Tubulin; C095114:collapsin response mediator protein-2; C060606:numb protein, Drosophila; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases