Multiple kinesins speed up cargo transport in crowded environments by sharing load.

Kinesin motors transport cargoes along microtubules inside of cells. Although it is well known that the cargoes are typically carried by multiple kinesins and that the more motors used, the further the cargoes travel, it has been challenging to determine the number of motors moving a cargo and any instant. Further, there is no unified statement on the relationship between cargo velocity and motor number, especially in the presence of a very crowded cytoplasmic environment. Here, we use a non-invasive method to quantify instantaneous motor number, and use it to investigate the effects of crowded environments on cargo motion when it is carried by multiple kinesins. Our experiments reveal that the velocity of the cargo depends on the number of motors on the cargo and the size of the crowders in crowded environments. Our finding suggests that kinesin tension plays a role in collective motion, which has been confirmed through stochastic kinesin simulations. Overall, our study demonstrates the broad applicability of the non-invasive method to determine engaged motor numbers and sheds light on the intriguing interplay between macromolecular crowding, kinesin tension, and kinesin-mediated cargo transport.