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Cell and Tissue Research

, Volume 321, Issue 1, pp 97–104 | Cite as

Nuclear localization of the major vault protein in U373 cells

  • Marco Slesina
  • Elisabeth M. Inman
  • Leonard H. Rome
  • Walter VolknandtEmail author
Regular Article

Abstract

The major vault protein (MVP) is the predominant member of a large ribonucleoprotein particle, named vault. Vaults are abundant in the cytosol of mammalian cells. Mammalian MVP has previously been reported to be associated with the nucleus, particularly its cytosolic surface on which vaults are thought to dock at or near the nuclear pore complex. To date the presence of vault particles inside the nucleus has been convincingly reported only for sea urchin cells. We have addressed the potential nuclear localization of MVP in mammalian cells by employing confocal laser microscopy and cryo-immunoelectron microscopy. As revealed by immunostaining and by analysis of cells transfected with a construct encoding MVP and green fluorescent protein, MVP is present in both the cytosol and in the nucleus. Cryo-electron microscopy of human astroglioma U373 cells reveals clusters of immunogold particles at nuclear pores and in the nucleoplasm suggesting that nuclear MVP is associated with particulate structures. Quantification of the fluorescence observed in the cytosol and in the nuclei reveals that about 5% of the MVP in U373 cells is localized inside the nucleus. Our results further support the notion that part of the cellular MVP can enter the nucleus.

Keywords

Major vault protein Nuclear pore complex Ribonucleoprotein particles Vaults Human astrogliomal U373 cells Rat pheochromocytoma cell line 

Notes

Acknowledgements

We are grateful to Dr. Herbert Zimmermann for valuable suggestions and for critically reading the manuscript. We are also grateful to Dr. Valerie Kickhoefer for support and for supplying us with antibodies.

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Marco Slesina
    • 1
  • Elisabeth M. Inman
    • 2
    • 3
  • Leonard H. Rome
    • 2
  • Walter Volknandt
    • 1
    Email author
  1. 1.Zoological Institute, AK NeurochemistryBiocenter J.W. Goethe UniversityFrankfurt am MainGermany
  2. 2.Department of Biological ChemistryUCLA School of Medicine and the Jonsson Comprehensive Cancer CenterLos AngelesUSA
  3. 3.Wound Management R&D, Hyland Immuno DivisionBaxter Healthcare CorporationDuarteUSA

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