Abstract
Microtubules and actin filaments function coordinately in many cellular processes1–3. Although much of this coordination is mediated by proteins that statically bridge the two cytoskeletal networks4–6, kinesin-14 motors with an actin binding calponin homology domain (KCHs) have been discovered as putatively dynamic crosslinkers in plants7,8. OsKCH1, a KCH from rice, interacts with both microtubules and actin filaments in vivo and in vitro9. However, it has remained unclear whether this interaction is dynamic or if actin binding reduces or even abolishes the motor's motility on microtubules10,11. Here, we directly show in vitro that OsKCH1 is a non-processive, minus-end-directed motor that transports actin filaments along microtubules. Interestingly, we observe two distinct transport velocities dependent on the relative orientation of the actin filaments with respect to the microtubules. In addition, torsional compliance measurements on individual molecules reveal low flexibility in OsKCH1. We suggest that the orientation-dependent transport velocities emerge from OsKCH1's low torsional compliance combined with an inherently oriented binding to the actin filament. Together, our results imply a central role of OsKCH1 in the polar orientation of actin filaments along microtubules, and thus a contribution to the organization of the cytoskeletal architecture.
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Acknowledgements
We thank C. Thodte, C. Bräuer and J. Sandberg-Meinhardt for technical assistance, P. Nick (KIT) for providing the OsKCH(aa1-744) expression plasmid and T. Scholz (Hannover Medical School) for providing rabbit muscle actin. W.J.W. and S.D. acknowledge support from the European Research Council (ERC starting grant 242933 to S.D.) and the Deutsche Forschungsgemeinschaft (Heisenberg programme grant DI 1226/4 and research unit SFG 877 grant DI 1226/5).
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W.J.W. and S.D. initiated research, designed experiments and wrote the manuscript. W.J.W. created reagents. W.J.W., I.M. and F.R. performed experiments and analysed data. All authors read the manuscript.
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Walter, W., Machens, I., Rafieian, F. et al. The non-processive rice kinesin-14 OsKCH1 transports actin filaments along microtubules with two distinct velocities. Nature Plants 1, 15111 (2015). https://doi.org/10.1038/nplants.2015.111
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DOI: https://doi.org/10.1038/nplants.2015.111
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