Spinal Muscular Atrophy: SMNs role in MRNA localization and local translation

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DESCRIPTION (provided by applicant): Spinal Muscular Atrophy (SMA) is a common inherited neurodegenerative disease caused by mutations or deletions in the SMN1 gene that encodes for the ubiquitous Survival of Motor Neuron Protein (SMN), which is essential for the assembly of spliceosomal snRNP complexes in all cells. A major unanswered question is how the loss of SMN leads to neuronal dysfunction and SMA. Further work is greatly needed to identify functions for SMN in neurons. Immunohistochemical studies have revealed localization of SMN to axons and dendrites from sections of spinal cord. Using high-resolution immunofluorescence of cultured neurons, we have shown that SMN forms granules which colocalize with RNA and ribosomes in growth cones. Using live cell imaging, we showed that EGFP-SMN granules exhibited rapid and bi-directional movements in neuronal processes and growth cones of live neurons. Primary motor neurons cultured from a transgenic mouse model of SMA displayed axonal defects that include loss of beta-actin mRNA from growth cones, suggesting a novel function for SMN in the mechanism of mRNA localization. Our preliminary data indicate that SMN associates with the mRNA binding proteins, ZBP1, ZBP2 and HuD, which are known to be involved in neuronal mRNA localization and stability. We hypothesize that these mRNA binding proteins interact with SMN to facilitate mRNA localization in processes and stabilization within growth cones. Experiments proposed here will further characterize the association, molecular interactions and dynamic regulation between SMN, HuD, ZBPs and associated mRNAs within RNA transport granules. SMN deficient motor neurons will be used to identify specific mRNAs that are altered in their localization and translation. We will investigate whether dysfunction of local mRNA regulation within SMN-deficient neurons contributes to altered growth cone motility. This research will provide new insight into neuronal functions for SMN, whereby possible defects in the assembly, localization and/or translation of mRNP complexes may contribute to the disease process in SMA.