UNC-104/KIF1A is a Kinesin-3 motor that transports synaptic vesicles from the cell body towards the synapse by binding to PI(4,5)P₂ through its PH domain. The fate of the motor upon reaching the synapse is not known. We found that wild-type UNC-104 is degraded at synaptic regions through the ubiquitin pathway and is not retrogradely transported back to the cell body. As a possible means to regulate the motor, we tested the effect of cargo binding on UNC-104 levels. The unc-104(e1265) allele carries a point mutation (D1497N) in the PI(4,5)P₂ binding pocket of the PH domain, resulting in greatly reduced preferential binding to PI(4,5)P₂ in vitro and presence of very few motors on pre-synaptic vesicles in vivo. unc-104(e1265) animals have poor locomotion irrespective of in vivo PI(4,5)P₂ levels due to reduced anterograde transport. Moreover, they show highly reduced levels of UNC-104 in vivo. To confirm that loss of cargo binding specificity reduces motor levels, we isolated two intragenic suppressors with compensatory mutations within the PH domain. These show partial restoration of in vitro preferential PI(4,5)P₂ binding and presence of more motors on pre-synaptic vesicles in vivo. These animals show improved locomotion dependent on in vivo PI(4,5)P₂ levels, increased anterograde transport, and partial restoration of UNC-104 protein levels in vivo. For further proof, we mutated a conserved residue in one suppressor background. The PH domain in this triple mutant lacked in vitro PI(4,5)P₂ binding specificity, and the animals again showed locomotory defects and reduced motor levels. All allelic variants show increased UNC-104 levels upon blocking the ubiquitin pathway. These data show that inability to bind cargo can target motors for degradation. In view of the observed degradation of the motor in synaptic regions, this further suggests that UNC-104 may get degraded at synapses upon release of cargo. The cell body and the synapse in a neuron are often separated by significant distance, which is spanned by the axon connecting the two. Transport of various cargoes along the axonal highway is very important for neuronal function. The regulation of this complex process is not well understood. Using the Caenorhabditis elegans model system, we have demonstrated for the first time the fate of a motor after it carries its cargo to the synapse from the cell body. We show that the UNC-104 motor, which carries pre-synaptic vesicles to the synapse, is degraded once it gets there. Moreover, our genetic studies show evidence that loss of cargo binding targets the motor for degradation, suggesting an attractive mechanism for the regulation of motors at the synapse. Our study opens up several further questions, such as the mechanism of motor degradation, and has significant implications for regulation of cargo transport.

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