Nogo-A inhibits necdin-accelerated neurite outgrowth by retaining necdin in the cytoplasm.
Nogo-A has been identified in the central nervous system as an inhibitor for axonal regeneration. Previous works have mainly focused on Nogo-A in oligodendrocytes and the roles of neuronal intracellular Nogo-A remain elusive. To gain deep insight into the physiological functions of Nogo-A, a yeast two-hybrid screening was performed with ... Nogo-66 as bait. We identified a new interaction between Nogo-66 and necdin. Mutagenesis analysis revealed that the central region of necdin was indispensable for the interaction of necdin with Nogo-66. The interaction was further confirmed by co-immunoprecipitation in neural tissues and cultured cortical neurons. Morphological evidence showed that Nogo-A and necdin highly colocalized in rat cortical and dorsal root ganglia neurons. Ectopic expression of Nogo-A in HEK293 cells led to retention of necdin from the nucleus to the cytoplasm. Furthermore, overexpression of Nogo-A in PC12 cells and cultured cortical neurons inhibited necdin-accelerated neurite outgrowth. Meanwhile, necdin was found to be significantly sequestered in the cytoplasm of PC12 cells stably overexpressing Nogo-A. Together, these data suggest that Nogo-A is a novel necdin binding protein and inhibits necdin-accelerated neuronal neurite outgrowth by sequestering necdin in the cytoplasm.
Mesh Terms:
Amino Acid Sequence, Animals, Cells, Cultured, Cytoplasm, Humans, Male, Molecular Sequence Data, Myelin Proteins, Nerve Regeneration, Nerve Tissue Proteins, Neurites, Neurons, Nuclear Proteins, PC12 Cells, Rats, Rats, Sprague-Dawley, Sequence Alignment, Two-Hybrid System Techniques
Amino Acid Sequence, Animals, Cells, Cultured, Cytoplasm, Humans, Male, Molecular Sequence Data, Myelin Proteins, Nerve Regeneration, Nerve Tissue Proteins, Neurites, Neurons, Nuclear Proteins, PC12 Cells, Rats, Rats, Sprague-Dawley, Sequence Alignment, Two-Hybrid System Techniques
Mol. Cell. Neurosci.
Date: May. 01, 2009
PubMed ID: 19386232
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