S-Acylation of Neuronal Transport Machinery: A Focused Exploration of p150Glued S-Acylation in Neurons

ABSTRACT

Neurons require efficient trafficking and delivery of neuronal proteins and organelles to specific subcellular locations. Dynein and kinesin motors mediate long-distance transport of cargo along microtubules spanning the axon to maintain neuronal function. The activity of motor proteins is tightly regulated, and aberrant activity can result in various neuropathies. S-acylation is an important mechanism to regulate neuronal protein trafficking. I identified several motor protein subunits and their activators in S-acyl-proteomic studies as likely to be S-acylated. The Sanders Lab recently confirmed S-acylation of the dynein activating complex dynactin subunit p150Glued. p150Glued is the largest dynactin subunit and mediates dynein complex microtubule binding and processive motility. Due to the importance of p150Glued in dynein-mediated axonal transport, I hypothesized that the functional role of p150Glued S-acylation is to regulate this trafficking process. To investigate this, I characterized p150Glued S-acylation during hippocampal culture development and identified a peak of S-acylation around day in vitro 14 and sought to determine if p150Glued S-acylation is reversible by analyzing turnover of p150Glued S-acylation and protein. I also developed neuronal culture models to replace endogenous p150Glued with GFP-tagged wild type or S-acylation deficient variants and to confirm the identity of the p150Glued S-acylating enzyme.