title: ZDHHC3 Tyrosine Phosphorylation Regulates Neural Cell Adhesion Molecule Palmitoylation.
abstract: The neural cell adhesion molecule (NCAM) mediates cell-cell and cell-matrix adhesion. It is broadly expressed in the nervous system and regulates neurite outgrowth, synaptogenesis, and synaptic plasticity. Previous in vitro studies revealed that palmitoylation of NCAM is required for fibroblast growth factor 2 (FGF2)-stimulated neurite outgrowth and identified the zinc finger DHHC (Asp-His-His-Cys)-containing proteins ZDHHC3 and ZDHHC7 as specific NCAM-palmitoylating enzymes. Here, we verified that FGF2 controlled NCAM palmitoylation in vivo and investigated molecular mechanisms regulating NCAM palmitoylation by ZDHHC3. Experiments with overexpression and pharmacological inhibition of FGF receptor (FGFR) and Src revealed that these kinases control tyrosine phosphorylation of ZDHHC3 and that ZDHHC3 is phosphorylated by endogenously expressed FGFR and Src proteins. By site-directed mutagenesis, we found that Tyr18 is an FGFR1-specific ZDHHC3 phosphorylation site, while Tyr295 and Tyr297 are specifically phosphorylated by Src kinase in cell-based and cell-free assays. Abrogation of tyrosine phosphorylation increased ZDHHC3 autopalmitoylation, enhanced interaction with NCAM, and upregulated NCAM palmitoylation. Expression of ZDHHC3 with tyrosine mutated in cultured hippocampal neurons promoted neurite outgrowth. Our findings for the first time highlight that FGFR- and Src-mediated tyrosine phosphorylation of ZDHHC3 modulates ZDHHC3 enzymatic activity and plays a role in neuronal morphogenesis.
authors.affiliation: Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, Genoa, Italy Department of Neurosciences, Biomedicine and Movement Sciences, Section of Biology and Genetics, University of Verona, Verona, Italy.
authors.forename: Patricia Marie-Jeanne
authors.initials: PM
authors.lastname: Lievens
authors.affiliation: Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, Genoa, Italy St. Petersburg State University, St. Petersburg, Russia.
authors.forename: Tatiana
authors.initials: T
authors.lastname: Kuznetsova
authors.affiliation: Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, Genoa, Italy.
authors.forename: Gaga
authors.initials: G
authors.lastname: Kochlamazashvili
authors.affiliation: Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, Genoa, Italy.
authors.forename: Fabrizia
authors.initials: F
authors.lastname: Cesca
authors.affiliation: Department of Cellular Neurophysiology, Hannover Medical School, Hannover, Germany.
authors.forename: Natalya
authors.initials: N
authors.lastname: Gorinski
authors.affiliation: Department of Cellular Neurophysiology, Hannover Medical School, Hannover, Germany.
authors.forename: Dalia Abdel
authors.initials: DA
authors.lastname: Galil
authors.affiliation: Department of Cellular Neurophysiology, Hannover Medical School, Hannover, Germany.
authors.forename: Volodimir
authors.initials: V
authors.lastname: Cherkas
authors.affiliation: Department of Physiological Chemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Natalia
authors.initials: N
authors.lastname: Ronkina
authors.affiliation: Department of Physiological Chemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Juri
authors.initials: J
authors.lastname: Lafera
authors.affiliation: Department of Physiological Chemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Gaestel
authors.affiliation: Department of Cellular Neurophysiology, Hannover Medical School, Hannover, Germany ponimaskin.evgeni@mh-hannover.de alexander.dityatev@dzne.de.
authors.forename: Evgeni
authors.initials: E
authors.lastname: Ponimaskin
authors.affiliation: Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, Genoa, Italy Molecular Neuroplasticity Group, DZNE, Magdeburg, Germany Medical Faculty, Otto von Guericke University, Magdeburg, Germany Center for Behavioral Brain Sciences (CBBS), Magdeburg, Germany ponimaskin.evgeni@mh-hannover.de alexander.dityatev@dzne.de.
authors.forename: Alexander
authors.identifier: 0000-0002-0472-0553
authors.initials: A
authors.lastname: Dityatev
mesh_terms: D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D005786:Gene Expression Regulation; D054878:Lipoylation; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009154:Mutation; D019006:Neural Cell Adhesion Molecules / Q000378:metabolism*; D016501:Neurites / Q000378:metabolism; D010766:Phosphorylation; D017468:Receptors, Fibroblast Growth Factor / Q000378:metabolism*; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism*
chemical_list: C465718:GODZ protein, mouse; D008565:Membrane Proteins; D019006:Neural Cell Adhesion Molecules; D017468:Receptors, Fibroblast Growth Factor; D016222:Fibroblast Growth Factor 2; D014443:Tyrosine; D000217:Acyltransferases; D019061:src-Family Kinases