Optogenetic control of kinesin-1, -2, -3 and dynein reveals their specific roles in vesicular transport
Sahil Nagpal,
Karthikeyan Swaminathan,
Daniel Beaudet,
Maud Verdier,
Samuel Wang,
Christopher L. Berger,
Florian Berger,
Adam G. Hendricks
Affiliations
Sahil Nagpal
Department of Bioengineering, McGill University, Montreal, QC H3A 0E9, Canada
Karthikeyan Swaminathan
Department of Bioengineering, McGill University, Montreal, QC H3A 0E9, Canada
Daniel Beaudet
Department of Bioengineering, McGill University, Montreal, QC H3A 0E9, Canada
Maud Verdier
Department of Bioengineering, McGill University, Montreal, QC H3A 0E9, Canada; Department of Biomedical Engineering and Health, Episen, Université Paris-Est Créteil, 94010 Créteil Cedex, France
Samuel Wang
Department of Bioengineering, McGill University, Montreal, QC H3A 0E9, Canada
Christopher L. Berger
Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, VT 05405-0075, USA
Florian Berger
Cell Biology, Neurobiology, and Biophysics, Department of Biology, Utrecht University, Utrecht, the Netherlands
Adam G. Hendricks
Department of Bioengineering, McGill University, Montreal, QC H3A 0E9, Canada; Corresponding author
Summary: Each cargo in a cell employs a unique set of motor proteins for its transport. To dissect the roles of each type of motor, we developed optogenetic inhibitors of endogenous kinesin-1, -2, -3 and dynein motors and examined their effect on the transport of early endosomes, late endosomes, and lysosomes. While kinesin-1, -3, and dynein transport vesicles at all stages of endocytosis, kinesin-2 primarily drives late endosomes and lysosomes. Transient optogenetic inhibition of kinesin-1 or dynein causes both early and late endosomes to move more processively by relieving competition with opposing motors. Kinesin-2 and -3 support long-range transport, and optogenetic inhibition reduces the distances that their cargoes move. These results suggest that the directionality of transport is controlled through regulating kinesin-1 and dynein activity. On vesicles transported by several kinesin and dynein motors, modulating the activity of a single type of motor on the cargo is sufficient to direct motility.
