Enhanced processivity and collective force production of kinesin-1 at low radial forces

Kinesin-1 is a robust motor that carries intracellular cargos towards the plus ends of microtubules. However, optical trapping studies reported that kinesin-1 is a slippery motor that quickly detaches from the microtubule, and multiple kinesins are incapable of teaming up to generate large collective forces. This may be due to the vertical (z) forces that the motor experiences in a single bead trapping assay, accelerating the detachment of the motor from a microtubule. Here, we substantially lowered the z-force by using a long DNA handle between the motor and the trapped bead and characterized the motility and force generation of single and multiple kinesin-1s. Contrary to previous views, we show that kinesin-1 is a robust motor that resists microtubule detachment before it reaches high hindering forces, but it quickly detaches under assisting forces even at low z-forces. We also demonstrate highly efficient collective force generation by multiple kinesin-1 motors. These results provide..., , # Enhanced processivity and collective force production of kinesin-1 at low radial forces

Dataset DOI: [10.5061/dryad.44j0zpcvz](https://doi.org/10.5061/dryad.44j0zpcvz)

## Description of the data and file structure

**The data falls into 3 broad categories:**

Optical Trapping

TIRF Microscopy

Refeyn

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**Refeyn:**

The raw Refeyn data was taken on a TwoMP mass photometer, and the calibrated files has been exported in the .csv format.

**TIRF:**

The TIRF microscopy data covers three experiments. In the first, oligo-labeled FL KIF5B is mixed with a complementary fluorescent DNA oligo and allowed to walk on microtubules decorated with MAP7. The 488 channel is KIF5B-GFP, and the other channel is the Cy3 oligo. This data was used exclusively for illustrative purposes and was not analyzed.

In the second, FL KIF5B is attached to streptavidin quantum dots via a DNA handle and allowed to walk on axonemes decorated with MAP7. This assay was used as a control for the DNA handle attach...,