, 2009). Overall, these

studies suggest that the highly conserved CAP-Gly domain in dynactin might be fully dispensable for vesicular transport in the cell. Strikingly, however, genetic evidence reveals that the CAP-Gly domain of p150Glued is essential for normal neuronal function since point mutations within this domain cause two autosomal dominant human neurodegenerative disorders: Perry syndrome and distal hereditary motor neuropathy 7B (HMN7B, also known as distal spinal and bulbar muscular atrophy) (Farrer et al., 2009 and Puls et al., 2003). HMN7B is caused by a glycine to serine substitution at residue 59 (G59S), while Perry syndrome is caused by one of five point mutations at residues 71, 72, or 74 (G71R, learn more G71E, G71A, T72P, Q74P) (Figures 1A and 1A′; see Movie S1 available online). The neuronal populations that degenerate in these two diseases are wholly distinct.

HMN7B affects motor neurons, while Perry syndrome primarily affects dopaminergic neurons in the substantia nigra (Puls et al., 2005 and Wider and Wszolek, 2008). It remains entirely unclear how these mutations, only 12–15 amino acids apart, differentially Metabolism inhibitor disrupt CAP-Gly domain function causing two disparate diseases. Here, we report a specific function for the CAP-Gly domain of dynactin in neurons. Our data show that the CAP-Gly domain enhances the distal enrichment of dynactin in the neuron, leading to efficient flux of cargo from the distal neurite. This function is separable from the role of dynactin in promoting bidirectional transport along the axon. Further, we show that the known disease-associated mutations all affect CAP-Gly function but differentially affect dynein-mediated transport along the axon, leading to a potential mechanistic explanation for the differential cell-type-specific degeneration observed in HMN7B and Perry syndrome. Together, these studies establish a role for the highly conserved CAP-Gly domain of dynactin in the efficient initiation of transport in highly polarized cells. These findings therefore provide insight Parvulin into both the regulation of axonal transport in the neuron and the cellular

basis for the neuronal specificity of mutations in dynactin. Multiple splice forms of p150Glued are expressed in brain, including a neuronally enriched p135 isoform that lacks the CAP-Gly domain (Tokito et al., 1996). We asked which p150Glued isoforms are recruited to cargos actively transported through the cell. Quantitative analysis of the p150Glued isoforms that copurified with LAMP1-enriched lysosomal fractions indicated that the full-length polypeptide is preferentially enriched in this fraction (Figures 1B and 1C). As dynein drives the motility of lysosomes along axons (Hendricks et al., 2010), the enrichment of full-length p150Glued that we observe suggests that the CAP-Gly domain may serve a specific function in the active transport of these vesicles.