Active transport of the survival motor neuron protein and the role of exon-7 in cytoplasmic localization
ABSTRACT Spinal muscular atrophy (SMA) is a neurodegenerative disease caused by deletion and/or mutation of the survival motor neuron protein Gene (SMN1) that results in the expression of a truncated protein lacking the C terminal exon-7. Whereas SMN has been shown to be an important component of diverse ribonucleoprotein (RNP) complexes, its function in neurons is unknown. We hypothesize that the active transport of SMN may be important for neurite outgrowth and that disruption of exon-7 could impair its normal intracellular trafficking. SMN was localized in granules that were associated with cytoskeletal filament systems and distributed throughout neurites and growth cones. Live cell imaging of enhanced green fluorescent protein (EGFP)-SMN granules revealed rapid, bidirectional and cytoskeletal-dependent movements. Exon-7 was necessary for localization of SMN into the cytoplasm but was not sufficient for granule formation and transport. A cytoplasmic targeting signal within exon-7 was identified that could completely redistribute the nuclear protein D-box binding factor 1 into the cytoplasm. Neurons transfected with SMN lacking exon-7 had significantly shorter neurites, a defect that could be rescued by redirecting the exon-7 deletion mutant into neurites by a targeting sequence from growth-associated protein-43. These findings provide the first demonstration of cytoskeletal-based active transport of SMN in neuronal processes and the function of exon-7 in cytoplasmic localization. Such observations provide motivation to investigate possible transport defects or inefficiency of SMN associated RNPs in motor neuron axons in SMA.
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- "Nonetheless, the study provides unparalleled insights into the repertoire of GC RBPs. Out of the 22 putative RBPs identified, only two, zipcode binding protein 1 (ZBP1, also known as IMP-1 and Vg1RBP) and survival motor neuron 1 (SMN) have previously been identified in GCs (Zhang et al., 2001, 2003, 2006; Leung et al., 2006; Fallini et al., 2011; Welshhans and Bassell, 2011). The single largest group of RBPs, comprising about 50% of all RBPs identified in the GCs, were the heterogenous nuclear ribonucleoprotein family (hnRNP) family of RBPs, a large family of RBPs that varies greatly in both function and structure (Han et al., 2010). "
ABSTRACT: RNA localization and regulation play an important role in the developing and adult nervous system. In navigating axons, extrinsic cues can elicit rapid local protein synthesis that mediates directional or morphological responses. The mRNA repertoire in axons is large and dynamically changing, yet studies suggest that only a subset of these mRNAs are translated after cue stimulation, suggesting the need for a high level of translational regulation. Here, we review the role of RNA-binding proteins (RBPs) as local regulators of translation in developing axons. We focus on their role in growth, guidance, and synapse formation, and discuss the mechanisms by which they regulate translation in axons.Frontiers in Neuroscience 05/2013; 7:81. DOI:10.3389/fnins.2013.00081 · 3.70 Impact Factor
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- "Because global inhibition of granule secretion decreased mSmn levels in neurites, we further investigated whether this may lead to similar defects caused by SMN deficiency in NSC34D cells. SMN granules have been implicated in β-actin transport –. β-Actin is enriched in the growth cone and therefore mSmn-deficient motor neurons may be expected to exhibit defects in β-actin transport and cause growth cone defects . The growth cone was examined by staining for F-actin (red) and βIII-tubulin (white) followed by confocal microscopy (Figure 5A). "
ABSTRACT: Proximal spinal muscular atrophy (SMA) is a neurodegenerative disorder caused by deficiency of the ubiquitous Survival of Motor Neuron (SMN) protein. SMN has been shown to be transported in granules along the axon and moved through cytoskeletal elements. However, the role and nature of SMN granules are still not well characterized. Here, using immunocytochemical methods and time-lapse studies we show that SMN granules colocalize with the Golgi apparatus in motor neuron-like NSC34 cells. Electron microscopy clearly revealed that SMN granules are transported into the Golgi stack and aggregate in the trans-Golgi apparatus. SMN granules are characterized as either coated or un-coated and behave like regulated secretory granules. Treatment of cells with monensin to disrupt Golgi-mediated granule secretion decreased SMN expression in neurites and caused growth cone defects similar to those seen in SMN knockdown cells. Knockdown of Cop-α, the protein that coats vesicles transporting proteins between the Golgi compartments, caused SMN granule accumulation in the Golgi apparatus. In addition to the well-studied role of SMN in small nuclear ribonucleoprotein (SnRNP) assembly, this work links SMN granules with the Golgi network and thus sheds light on Golgi-mediated SMN granule transport.PLoS ONE 12/2012; 7(12):e51826. DOI:10.1371/journal.pone.0051826 · 3.23 Impact Factor
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- "Thus, the SMN YG-box requires at least three residues following conserved Gly279 for efficient oligomerization, with no apparent consequences when either of the three is substituted by alanine in the context of full-length exon 7. For correct cellular localization and function, however, it appears that at least seven residues following Gly279 are required, including the moderately well-conserved QNQKE motif starting at Gln282(Carrel et al., 2006; Hua and Zhou, 2004; Zhang et al., 2003). "
ABSTRACT: The survival motor neuron (SMN) protein forms the oligomeric core of a multiprotein complex that functions in spliceosomal snRNP biogenesis. Loss of function mutations in the SMN gene cause spinal muscular atrophy (SMA), a leading genetic cause of infant mortality. Nearly half of the known SMA patient missense mutations map to the SMN YG-box, a highly conserved oligomerization domain of unknown structure that contains a (YxxG)(3) motif. Here, we report that the SMN YG-box forms helical oligomers similar to the glycine zippers found in transmembrane channel proteins. A network of tyrosine-glycine packing between helices drives formation of soluble YG-box oligomers, providing a structural basis for understanding SMN oligomerization and for relating defects in oligomerization to the mutations found in SMA patients. These results have important implications for advancing our understanding of SMN function and glycine zipper-mediated helix-helix interactions.Structure 09/2012; 20(11). DOI:10.1016/j.str.2012.08.024 · 6.79 Impact Factor