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Movement of vault particles visualized by GFP-tagged major vault protein

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Abstract

Vaults are abundant large ribonucleoprotein particles. They frequently colocalize with microtubules and accumulate in filamentous actin-rich lamellipodia. To examine the movement of vaults in living cells, a chimera between the green fluorescent protein and the major vault protein was created. This fusion protein assembled into vault particles as assayed by biochemical fractionation and direct observation of living or fixed cells. By fluorescence recovery after photobleaching, we analyzed the bulk transport of vault particles into neuritic tips of PC12 cells treated with nerve growth factor. Confocal laser scanning microscopy demonstrated co-localization of the major vault protein and microtubules. Video microscopy indicated that, whereas the majority of vault particles were stationary, some individual vault particles moved rapidly, consistent with the action of a microtubule-based or actin-based molecular motor.

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Acknowledgements

The authors thank Dr. Valerie Kickhoefer and Prof. Herbert Zimmermann for reading the manuscript and making valuable suggestions.

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Author notes

  1. Elisabeth M. Inman

    Present address: Wound Management R&D, Hyland Immuno Division, Baxter Healthcare Corporation, Duarte, CA, 91010, USA

Authors and Affiliations

  1. Biocenter, Zoological Institute, J. W. Goethe University, Marie-Curie-Strasse 9, 60439, Frankfurt, Main, Germany

    Marco Slesina & Walter Volknandt

  2. Department of Biological Chemistry, UCLA School of Medicine and the Jonsson Comprehensive Cancer Center, Los Angeles, CA, 90095-1737, USA

    Elisabeth M. Inman, Ann E. Moore & Leonard H. Rome

  3. Department of Medicine/Cardiology, UCLA School of Medicine, Los Angeles, CA, 90095-1737, USA

    Joshua I. Goldhaber

Authors
  1. Marco Slesina
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  2. Elisabeth M. Inman
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  3. Ann E. Moore
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  4. Joshua I. Goldhaber
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  5. Leonard H. Rome
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  6. Walter Volknandt
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Corresponding author

Correspondence to Walter Volknandt.

Additional information

This work was supported by the United States Public Health Service, National Institutes of Health (grant GM38097 to L.H.R.) and by the North Atlantic Treaty Organization (grant CRG972834 to W.V.).

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Slesina, M., Inman, E.M., Moore, A.E. et al. Movement of vault particles visualized by GFP-tagged major vault protein. Cell Tissue Res 324, 403–410 (2006). https://doi.org/10.1007/s00441-006-0158-8

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  • DOI: https://doi.org/10.1007/s00441-006-0158-8

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