Bidirectional helical motility of cytoplasmic dynein around microtubules
- University of California, Berkeley, United States
Figures
Figure 1 with 1 supplement
3D tracking of dynein-driven transport along MT.
(A) Schematic representation of the experimental geometry (not to scale). The MT suspended bridge is formed by attaching an MT (green) to the surface-immobilized beads (gray) that are coated with a …
Figure 1—figure supplement 1
Kinesin-1 follows a single protofilament on a MT track.
(A) Example trace of a kinesin-1 coated bead, traveling along GMP-CPP MT filaments with a left handed spiraling motion. (B) Representative two dimensional trace. The average pitch of helical motion …
Figure 2 with 5 supplements
Dynein moves in both left- and right-handed helical paths along MT bridges.
(A and B) (top) Representative three-dimensional trace of a cargo bead-driven by GST-Dyn331kD motors shows left- (A) and right-handed (B) helical motion. (bottom) Two-dimensional projections of the …
Figure 2—figure supplement 1
Movement of a GST-Dyn331kD coated bead along an MT bridge.
(A) First 20 s of the 3D trace shown in Figure 2A is plotted in x, y, and z directions as a function of time. (B) The brightfield image of the bead shows the changes in the position and intensity of …
Figure 2—figure supplement 2
Additional example of the right-handed helical movement of a GST-Dyn331kD-coated bead along an MT bridge.
(top) Representative three-dimensional trace of a cargo bead-driven by GST-Dyn331kD motors shows right-handed helical motion. (bottom) Two-dimensional projection of the trace shown at top.
Figure 2—figure supplement 3
Bidirectional helical motility of cargo beads driven by full-length dynein along MT bridges.
(A and B) (top) Representative three-dimensional trace of a cargo bead driven by full-length dynein motors shows left- (A) and right-handed (B) helical motion. (bottom) Two-dimensional projections …
Figure 2—figure supplement 4
Change in handedness of rotation during the transport of a cargo bead driven by full-length dynein motors.
An example trace shows that a cargo bead initially moves along a GMP-CPP MT bridge with a right-handed helical motion. At around t = 5 s, the bead reverses its helical directionality (arrow) and …
Figure 2—figure supplement 5
GST-Dyn331kD on taxol stabilized MTs.
(A) Representative two-dimensional trace of cargo beads driven by GST-Dyn331kD on taxol stabilized MTs which contains the mixture of 12, 13, 14 protofilaments with different rotational pitches. (B) …
Figure 3
Single dynein motors frequently switch the direction of their sideways movement.
(A) Schematic representation of quantum-dot labeled single dynein motors on the MT bridges (not to scale). Expected amplitude of rotations is ∼50 nm. (B) Two example traces show 2D projection of …
Figure 4
Dynein monomers prefer to move in a right-handed helix.
(A) Representative three-dimensional trace of a cargo bead driven by monomers shows right-handed helical motion. (B) (top) Representative two-dimensional trace for monomeric Dyn331kD. (bottom) …
Figure 5
A model for the helical movement of cytoplasmic dynein.
(A) Top view of a monomeric dynein (red oval) stepping toward the MT-minus end (arrows). The yellow circles represent the putative binding sites for the highlighted dyneins. The closest available …
Author response image 1
Videos
Video 1
Example recording of left-handed rotations of a kinesin-1 coated bead at 100 ms time resolution via bright-field microscopy. Fluorescent image of MT has been superimposed onto the bright-field image for illustration purposes.
https://doi.org/10.7554/eLife.03205.005
Video 2
Example recording of left-handed rotations of GST-Dyn331kD-coated bead at 100 ms time resolution via bright-field microscopy.
https://doi.org/10.7554/eLife.03205.012
Video 3
Example recording of right-handed rotations of GST-Dyn331kD-coated bead at 100 ms time resolution via bright-field microscopy.
https://doi.org/10.7554/eLife.03205.013
Video 4
Example recording of right- and left-handed rotations of full-length dynein-coated bead at 100 ms time resolution via bright-field microscopy.
https://doi.org/10.7554/eLife.03205.014
Video 5
Example recording of right-handed rotations of monomeric Dyn331kD-coated bead. Video recorded via bright-field microscopy with time resolution of 100 ms.
https://doi.org/10.7554/eLife.03205.017Additional files
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Supplementary file 1
Matlab code used in data analysis.
- https://doi.org/10.7554/eLife.03205.019
