The Kinesin-12 Kif15 is a processive track-switching tetramer
- University of Warwick, United Kingdom
- Cancer Research UK, London Research Institute, United Kingdom
Figures
Figure 1
Kif15 is a tetramer.
(A) Coomassie stained SDS-PAGE gel of purified His6-hTpx2, His6-hKif15, and His6-hKif15-GFP. (B) Tetrameric His6-hKif15 on a 4–16% Native-PAGE gel stained with coomassie. Calculated molecular …
Figure 2 with 2 supplements
hKif15 is a processive tetramer.
(A) Kymographs showing typical behaviour of eGFP-labelled hKif15 motors on GMP-CPP stabilised microtubules (processive movement–P, diffusion–D, plus-end dwelling–asterix, processive minus-end …
Figure 2—figure supplement 1
TIRF-based analysis of hKif15 motility.
(A) Kymographs showing the behaviour of hKif15-eGFP motors on GMP-CPP stabilised microtubules in dependency indicated salt concentrations apart from the standard condition used throughout the …
Figure 2—figure supplement 2
Analysis of bead motility and microtubule-binding in laser trap experiments.
(A) Poisson statistics for the bead-binding probability of n ≥ 1 (blue) and n ≥ 2 (orange) motors fitted to data from a hKif15 dilution series (brown). Number of analysed beads per concentration >50 …
Figure 3
hKif15 effectively switches microtubules at intersections.
(A) Kymographs showing the processive switch of an eGFP-labelled hKif15 motor at a microtubule–microtubule intersection. The image in the upper left gives an overview of the microtubule positons, bar…
Figure 4
hKif15-eGFP can transport short microtubules as a cargo.
Kymographs showing examples of processive (left) and tug-of-war-type transport (right), with an overview image of the crosslinked microtubules on top. Left: hKif15-eGFP motors drive slow (26 nm•s−1) …
Figure 5
hTpx2 inhibits hKif15 motility by increasing its microtubule affinity.
(A) Formation of a stable hKif15/hTpx2 complex occurs only at low µM concentration of Tpx2. Coomassie stained SDS-PAGE gels showing the first 23 of 25 fractions of a 5–40% glycerol gradient in 35 mM …
Figure 6
Schematic model summarising the biophysical and biochemical properties of hKif15 and illustrating how the motor may operate within a k-fibre (parallel microtubule bundle).
A–hKif15 can move uni-directionally towards the plus-end (minus-end-directed motion can also occur albeit with lower frequency) by processive stepping or by bi-directionally diffusion along the …
Videos
Video 1
hKif15 effectively switches microtubules at intersections.
Video of the events summarised in Figure 3A (hKif15-eGFP—cyan, microtubules—yellow). The switching hKif15-eGFP motor is marked by the cyan arrowhead.
Video 2
hKif15 effectively switches microtubules at intersections.
Video of another hKif15-eGFP motor switching polarity-marked microtubules (hKif15-eGFP—cyan, seed—red, microtubule extension—yellow). The switching motor is marked by the cyan arrowhead. Please note …
Video 3
hKif15 effectively switches microtubules at intersections.
Video of the events summarised in Figure 3C (hKif15-eGFP—cyan, seed—red, microtubule extension—yellow). The switching hKif15-eGFP motor is marked by the cyan arrowhead.
Video 4
Surface-bound hKif15 does not support continuous sliding of GMP-CPP stabilised microtubules (white) over long distances.
https://doi.org/10.7554/eLife.01724.012
Video 5
