Inducible manipulation of motor-cargo interaction using engineered kinesin motors

Molecular motors drive long-range intracellular transport of various vesicles and other cargoes within a cell. Identifying which kinesin motors interact with which type of transport vesicles has been challenging, especially in complex neuronal cells. Here, we present a highly adaptable toolbox of engineered kinesin motors to control and interrogate the selectivity and regulation of cargo transport with acute chemical induction. Selectivity of cargo-motor interaction can be addressed by systematic screening of a library of kinesin tails and neuronal cargoes. Additionally, our toolbox can be used to study kinesin-cargo regulatory mechanisms, and we found that cargo trafficking by KIF16B is regulated by its PX domain. Furthermore, our toolbox enables acute manipulation of polarized trafficking in living neurons by steering transport into axons or dendrites. Engineering kinesin motors provides a powerful tool to map the specificity of interactions between kinesin and cargoes, manipulate polarized transport and investigate cargo-motor interaction modes.

Automated summary

The study introduces an engineered kinesin motor toolbox for exploring the selectivity and regulatory mechanisms of intracellular cargo transport. Systematic screening of different kinesin tails and neuronal vesicles can address the selectivity of cargo-motor interaction, and the toolbox allows for investigation of regulatory mechanisms of kinesin-cargo interactions, such as the PX domain regulation of cargo trafficking by KIF16B.