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Spatiotemporal control of microtubule nucleation and assembly using magnetic nanoparticles

Nature Nanotechnology volume 8, pages 199–205 (2013)Cite this article

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Abstract

Decisions on the fate of cells and their functions are dictated by the spatiotemporal dynamics of molecular signalling networks. However, techniques to examine the dynamics of these intracellular processes remain limited. Here, we show that magnetic nanoparticles conjugated with key regulatory proteins can artificially control, in time and space, the Ran/RCC1 signalling pathway that regulates the cell cytoskeleton. In the presence of a magnetic field, RanGTP proteins conjugated to superparamagnetic nanoparticles can induce microtubule fibres to assemble into asymmetric arrays of polarized fibres in Xenopus laevis egg extracts. The orientation of the fibres is dictated by the direction of the magnetic force. When we locally concentrated nanoparticles conjugated with the upstream guanine nucleotide exchange factor RCC1, the assembly of microtubule fibres could be induced over a greater range of distances than RanGTP particles. The method shows how bioactive nanoparticles can be used to engineer signalling networks and spatial self-organization inside a cell environment.

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Figure 1: Magnetic nanoparticles bioconjugated to signalling proteins controlling microtubule assembly.
Figure 2: Signalling proteins conjugated to magnetic nanoparticles promote microtubule assembly.
Figure 3: Magnetic control of microtubule nucleation and assembly.
Figure 4: Magnetically induced microtubule arrays are polarized, organized by dynein motors and can be manipulated by magnetic forces.
Figure 5: Microtubule assembly triggered by guanine exchange factor RCC1.

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Acknowledgements

The authors thank I. Mattaj and B. Koch for the RanQ69L and RCC1 expression plasmids, K. Weis for the Rango expression plasmid and N. Morin for the Ran(T24N) expression plasmid. The authors also thank F. Guyot, J.M. Guignier and L. Largeau for electron microscopy observations. Thanks also go to F. Grasset, A. Gautier, L. Jullien, T. Le Saux, A. Libchaber, N. Nerambourg, C. Tribet, J. Wu and the members of the Biophysical Chemistry Center of ENS for fruitful discussions. This work was supported by the CNRS, the Association pour la Recherche sur le Cancer (SFI20101201426), the Ligue Nationale Contre le Cancer (LNCC, 2009) and Ville de Paris ‘Emergence’ (to Z.G.), the Agence Nationale de la Recherche (ANR, ANR-08-PNANO-050 to R.LB., V.M., C.G. and Z.G.), FRM, Triangle de la Physique (to E.M.) and ATIP CNRS (to R.LB.).

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Authors and Affiliations

  1. Département de Chimie, Ecole Normale Supérieure, UMR 8640 CNRS-ENS-UPMC Pasteur, 24, rue Lhomond, Paris, 75005, France

    Céline Hoffmann & Zoher Gueroui

  2. Laboratoire de Photonique et de Nanostructures, LPN-CNRS, route de Nozay, Marcoussis, 91460, France

    Elsa Mazari & Charlie Gosse

  3. CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, F-35043, France

    Sylvie Lallet & Roland Le Borgne

  4. Université Rennes 1, UEB, IFR 140, Faculté de Médecine, Rennes, F-35043, France

    Roland Le Borgne

  5. CNRS UMR 6226, Université Rennes 1, Sciences Chimiques de Rennes, Campus de Beaulieu, Rennes, F-35042, France

    Valérie Marchi

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  1. Céline Hoffmann
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Contributions

C.H. and Z.G. conceived and designed the experiments. C.H., S.L. and Z.G. performed the experiments. E.M. and C.G. performed the simulations. All authors analysed the data. C.G., R.L.B. and V.M. contributed materials/analysis tools. C.H. and Z.G. wrote the manuscript and all authors commented on it.

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Correspondence to Zoher Gueroui.

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Hoffmann, C., Mazari, E., Lallet, S. et al. Spatiotemporal control of microtubule nucleation and assembly using magnetic nanoparticles. Nature Nanotech 8, 199–205 (2013). https://doi.org/10.1038/nnano.2012.246

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