title: A CRM1-mediated nuclear export signal is essential for cytoplasmic localization of neurogenin 3 in neurons.
abstract: Neurogenin 3 (Ngn3), a proneural gene, regulates dendritogenesis and synaptogenesis in mouse hippocampal neurons. Ngn3 is transiently exported from the cell nucleus to the cytoplasm when neuronal polarity is initiated, suggesting that the nucleo-cytoplasmic transport of the protein is important for its action on neuronal development. In this study, we identified for the first time a functional nuclear export sequence (NES2; ¹³¹YIWALTQTLRIA¹⁴²) in Ngn3. The green fluorescent protein (EGFP)-NES2 fusion protein was localized in the cytoplasm and its nucleo-cytoplasmic shuttling was blocked by the CRM1 specific export inhibitor leptomycin B. Mutation of a leucine residue to alanine (L135A) in the NES2 motif resulted in both cytoplasmic and nuclear localization of the EGFP-NES2 fusion protein and in the nuclear accumulation of ectopic full-length myc-Ngn3. In addition, point mutation of the leucine 135 counteracted the effects of Ngn3 on neuronal morphology and synaptic inputs indicating that the cytoplasmic localization of Ngn3 is important for neuronal development. Pharmacological perturbation of the cytoskeleton revealed that cytoplasmic Ngn3 is associated with microtubules.
authors: Instituto Cajal, CSIC, Madrid, Spain.
authors: Julia
authors: J
authors: Simon-Areces
authors: Estefania
authors: E
authors: Acaz-Fonseca
authors: Isabel
authors: I
authors: Ruiz-Palmero
authors: Luis-Miguel
authors: LM
authors: Garcia-Segura
authors: Maria-Angeles
authors: MA
authors: Arevalo
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism*; D015153:Blotting, Western; D002467:Cell Nucleus / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D028884:Karyopherins / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000378:metabolism; D016296:Mutagenesis; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D049790:Nuclear Export Signals / Q000235:genetics*; D009841:Oligonucleotides / Q000235:genetics; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D014162:Transfection; D000097606:Exportin 1 Protein
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D017931:DNA Primers; D049452:Green Fluorescent Proteins; D028884:Karyopherins; D009419:Nerve Tissue Proteins; D049790:Nuclear Export Signals; D009841:Oligonucleotides; D018160:Receptors, Cytoplasmic and Nuclear; D000097606:Exportin 1 Protein; C407737:Neurog3 protein, mouse; C485184:enhanced green fluorescent protein
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