title: Thioredoxin mediates oxidation-dependent phosphorylation of CRMP2 and growth cone collapse.
abstract: Semaphorin3A (Sema3A) is a repulsive guidance molecule for axons, which acts by inducing growth cone collapse through phosphorylation of CRMP2 (collapsin response mediator protein 2). Here, we show a role for CRMP2 oxidation and thioredoxin (TRX) in the regulation of CRMP2 phosphorylation and growth cone collapse. Sema3A stimulation generated hydrogen peroxide (H2O2) through MICAL (molecule interacting with CasL) and oxidized CRMP2, enabling it to form a disulfide-linked homodimer through cysteine-504. Oxidized CRMP2 then formed a transient disulfide-linked complex with TRX, which stimulated CRMP2 phosphorylation by glycogen synthase kinase-3, leading to growth cone collapse. We also reconstituted oxidation-dependent phosphorylation of CRMP2 in vitro, using a limited set of purified proteins. Our results not only clarify the importance of H2O2 and CRMP2 oxidation in Sema3A-induced growth cone collapse but also indicate an unappreciated role for TRX in linking CRMP2 oxidation to phosphorylation.
authors.affiliation: Laboratory of Intracellular Signaling, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Akifumi
authors.initials: A
authors.lastname: Morinaka
authors.forename: Mayumi
authors.initials: M
authors.lastname: Yamada
authors.forename: Rurika
authors.initials: R
authors.lastname: Itofusa
authors.forename: Yosuke
authors.initials: Y
authors.lastname: Funato
authors.forename: Yuta
authors.initials: Y
authors.lastname: Yoshimura
authors.forename: Fumio
authors.initials: F
authors.lastname: Nakamura
authors.forename: Takeshi
authors.initials: T
authors.lastname: Yoshimura
authors.forename: Kozo
authors.initials: K
authors.lastname: Kaibuchi
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Goshima
authors.forename: Mikio
authors.initials: M
authors.lastname: Hoshino
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kamiguchi
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Miki
mesh_terms: D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002642:Chick Embryo; D002522:Chlorocebus aethiops; D005260:Female; D005727:Ganglia, Spinal / Q000196:embryology / Q000378:metabolism; D055785:Gene Knockdown Techniques; D020439:Growth Cones / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008840:Microfilament Proteins; D008869:Microtubule-Associated Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D041681:NIH 3T3 Cells; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D010766:Phosphorylation; D011247:Pregnancy; D034741:RNA, Small Interfering / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D040121:Semaphorin-3A / Q000378:metabolism; D015398:Signal Transduction; D013879:Thioredoxins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D008840:Microfilament Proteins; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; C488217:Sema3a protein, mouse; D040121:Semaphorin-3A; C095114:collapsin response mediator protein-2; D013879:Thioredoxins; D006861:Hydrogen Peroxide; C507541:Mical1 protein, mouse; D006899:Mixed Function Oxygenases