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Processive bidirectional motion of dynein–dynactin complexes in vitro

Nature Cell Biology volume 8, pages 562–570 (2006)Cite this article

Abstract

Cytoplasmic dynein is the primary molecular motor responsible for transport of vesicles, organelles, proteins and RNA cargoes from the periphery of the cell towards the nucleus along the microtubule cytoskeleton of eukaryotic cells. Dynactin, a large multi-subunit activator of dynein, docks cargo to the motor and may enhance dynein processivity. Here, we show that individual fluorescently labelled dynein–dynactin complexes exhibit bidirectional and processive motility towards both the plus and minus ends of microtubules. The dependence of this activity on substrate ATP concentration, nucleotide analogues and inhibitors suggests that bidirectional motility is an active energy-transduction property of dynein–dynactin motor mechano-chemistry. The unique motility characteristics observed may reflect the flexibility of the dynein structure that leads to an enhanced ability to navigate around obstacles in the cell.

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Figure 1: Purification and characterization of dynein–dynactin–GFP from GFP–dynamitin mouse brains.
Figure 2: Example traces of motion exhibited by dynein–dynactin–GFP as visualized by TIRF.
Figure 3: Histograms of velocities of individual dynein–dynactin–GFP complexes exhibiting processive runs.
Figure 4: ATP dependence of average velocity and the effects of dynein inhibitors.
Figure 5: Possible models for the bidirectional motility observed for dynein–dynactin.

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Acknowledgements

This work was supported by a National Institutes of Health (NIH) project program grant (P01–AR–051174) to the Pennsylvania Muscle Institute, by an ALS Association grant to E.L.F.H., and J.L.R. is supported by an NIH NRSA grant (1F32GM075754–01). The authors also thank A. Loh, R. Kudaravalli, H. Pham, W. Zhou and M. Tokito.

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  1. Department of Physiology and Pennsylvania Muscle Institute, University of Pennsylvania, Philadelphia, 19104, PA, USA

    Jennifer L. Ross, Karen Wallace, Henry Shuman, Yale E. Goldman & Erika L.F. Holzbaur

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Correspondence to Erika L.F. Holzbaur.

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Ross, J., Wallace, K., Shuman, H. et al. Processive bidirectional motion of dynein–dynactin complexes in vitro. Nat Cell Biol 8, 562–570 (2006). https://doi.org/10.1038/ncb1421

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