Abstract
Kinesins are microtubule-based motor proteins that transport cargo to specific locations within the cell. However, the mechanisms by which cargoes are directed to specific cellular locations have remained elusive. Here, we investigated the in vivo movement of the Schizosaccharomyces pombe kinesin Tea2 to establish how it is targeted to microtubule tips and cell ends. Tea2 is loaded onto microtubules in the middle of the cell, in close proximity to the nucleus, and then travels using its intrinsic motor activity primarily at the tips of polymerizing microtubules. The microtubule-associated protein Mal3, an EB1 homologue, is required for loading and/or processivity of Tea2 and this function can be substituted by human EB1. In addition, the cell-end marker Tea1 is required to anchor Tea2 to cell ends. Movement of Tea1 and the CLIP170 homologue Tip1 to cell ends is abolished in Tea2 rigor (ATPase) mutants. We propose that microtubule-based transport from the vicinity of the nucleus to cell ends can be precisely regulated, with Mal3 required for loading/processivity, Tea2 for movement and Tea1 for cell-end anchoring.
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Acknowledgements
The authors wish to thank T. Toda, T. Niccoli, K. Leonhard and S. Castagnetti for critical reading of the manuscript, members of Cancer Research UK light microscopy facility for help with image acquisition and analysis, members of the Cell Cycle lab for helpful discussions and suggestions, J. R. McIntosh and R. West (University of Colorado, Boulder) for fruitful discussions and critical reading of the manuscript, A. Decottignies and P. Zarzov for advice on construction of mutant alleles, U. Fleig for pREPEB1 (Heinrich-Heine-Universitat Dusseldorf) and N. Peat for help with the manuscript. This work was supported by Cancer Research UK (formerly the Imperial Cancer Research Fund), a postdoctoral fellowship to H. Browning from the International Agency for Research on Cancer, and grant NS28562 to D. Hackney.
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Browning, H., Hackney, D. & Nurse, P. Targeted movement of cell end factors in fission yeast. Nat Cell Biol 5, 812–818 (2003). https://doi.org/10.1038/ncb1034
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DOI: https://doi.org/10.1038/ncb1034
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