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


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.


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



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.


  1. Abbondanza C, Rossi V, Rossigno A, Gallo L, Belsito A, Piluso G, Medici N, Nigro V, Molinari AM, Moncharmant B, Puca GA (1998) Interaction of vault particles with estrogen receptor in the MCF-7 breast cancer cell. J Cell Biol 141:1301–1310CrossRefPubMedGoogle Scholar
  2. Berger W, Elbling L, Miksche M (2000) Expression of the major vault protein LRP in human non-small-cell lung cancer cells: activation by short-term exposure to antineoplastic drugs. Int J Cancer 88:293–300CrossRefPubMedGoogle Scholar
  3. Berger W, Spiegl-Kreinecker S, Buchrotthner J, Elbling L, Pirker C, Fischer J, Micksche M (2001) Overexpression of the human major vault protein in astrocytic brain tumor cells. Int J Cancer 94:377–382CrossRefPubMedGoogle Scholar
  4. Chugani DC, Rome LH, Kedersha NL (1993) Localization of vault particles to the nuclear pore complex. J Cell Sci 106:23–29PubMedGoogle Scholar
  5. Feldherr CM, Akin D, Cohen RJ (2001) Regulation of functional nuclear pore size in fibroblasts. J Cell Sci 114:4621–4627PubMedGoogle Scholar
  6. Hamill DR, Suprenant KA (1997) Characterization of the sea urchin major vault protein: a possible role for vault ribonucleoprotein particles in nucleocytoplasmic transport. Dev Biol 190:117–128CrossRefPubMedGoogle Scholar
  7. Herrmann C, Volknandt W, Wittich B, Kellner R, Zimmermann H (1996) The major vault protein (MVP100) is contained in cholinergic nerve terminals of electric ray electric organ. J Biol Chem 271:13908–13915CrossRefPubMedGoogle Scholar
  8. Herrmann C, Golkaramnay E, Inman E, Rome LH, Volknandt W (1999) Recombinant major vault protein is targeted to neuritic tips of PC12 cells. J Cell Biol 144:1163–1172CrossRefPubMedGoogle Scholar
  9. Izquierdo MA, Scheffer GL, Flens MJ, Shoemaker RLH, Scheper RJ (1996) Relationship of LRP-human major vault protein to in vitro and clinical resistance to anticancer drugs. Cytotechnology 19:191–197CrossRefPubMedGoogle Scholar
  10. Kedersha NL, Rome LH (1986) Isolation and characterization of a novel ribonucleoprotein particle: large structures contain a single species of small RNA. J Cell Biol 103:699–709CrossRefPubMedGoogle Scholar
  11. Kedersha NL, Miquel MC, Bittner D, Rome LH (1990) Vaults: II. Ribonucleoprotein structures are highly conserved among higher and lower eukaryotes. J Cell Biol 110:895–901CrossRefPubMedGoogle Scholar
  12. Kedersha NL, Heuser JE, Chugani DC, Rome LH (1991) Vaults: III. Vault ribonucleoprotein particles open into flower-like structures with octagonal symmetry. J Cell Biol 112:225–235CrossRefPubMedGoogle Scholar
  13. Kickhoefer V, Rome LH (1994) The sequence of a cDNA encoding the major vault protein from Rattus norvegicus.Gene 151:257-260CrossRefPubMedGoogle Scholar
  14. Kickhoefer VA, Searles RP, Kedersha NL, Garber ME, Johnson DL, Rome LH (1993) Vault RNP particles from rat and bullfrog contain a related small RNA that is transcribed by RNA polymerase III. J Biol Chem 268:7868–7873PubMedGoogle Scholar
  15. Kickhoefer VA, Vasu SK, Rome LH (1996) Vaults are the answer, what is the question? Trends Cell Biol 6:174–178CrossRefPubMedGoogle Scholar
  16. Kickhoefer VA, Rajavel KS, Scheffer GL, Dalton WS, Scheper RJ, Rome LH (1998) Vaults are up-regulated in multidrug-resistant cancer cell lines. J Biol Chem 273:8971–8974CrossRefPubMedGoogle Scholar
  17. Kickhoefer VA, Siva AC, Kedersha NL, Inman EM, Ruland C, Streuli M, Rome LH (1999a) The 193-kD vault protein, VPARP, is a novel poly(ADP-ribose) polymerase. J Cell Biol 146:917–928CrossRefPubMedGoogle Scholar
  18. Kickhoefer VA, Stephen AG, Harrington L, Robinson MO, Rome LH (1999b) Vaults and telomerase share a common subunit, TEP1. J Biol Chem 274:32712–32718CrossRefPubMedGoogle Scholar
  19. Kickhoefer VA, Poderycki MJ, Chan EKL, Rome LH (2002) The La RNA-binding protein interacts with the vault RNA and is a vault-associated protein. J Biol Chem 277:41282–41286CrossRefPubMedGoogle Scholar
  20. Kiseleva E, Goldberg MW, Daneholt B, Allen TD (1996) RNP export is mediated by structural rorganization of the nuclear pore basket. J Mol Biol 250:304–311CrossRefGoogle Scholar
  21. Kitazano M, Sumizawa T, Takebayashi Y, Chen ZS, Furukawa T, Nagayama S, Tani A, Takao S, Aikou T, Akiyama S (1999a) Multidrug resistance and the lung resitance-related protein in human colon carcinoma SW-620 cells. J Natl Cancer Inst 91:1647–1653CrossRefPubMedGoogle Scholar
  22. Kitazano M, Okumura H, Ikeda R, Sumizawa T, Furukawa T, Nagayama S, Seto K, Aikou T, Akiyama S (1999b) Reversal of LRP-associated resistance in colon carcinoma SW-620 cells. Int J Cancer 91:126–131CrossRefGoogle Scholar
  23. Kong LB, Siva AC, Rome LH, Stewart PL (1999) Structure of the vault, a ubiquitous cellular component. Structure 7:371–379CrossRefPubMedGoogle Scholar
  24. Kong LB, Siva AC, Kickhoefer VA, Rome LH, Stewart PL (2000) RNA location and modeling of a WD40 repeat domain within the vault. RNA 6:1–11CrossRefPubMedGoogle Scholar
  25. Panté N, Kann M (2002) Nuclear pore complex is able to transport macromolecules with diameters of about 39 nm. Mol Biol Cell 13:425–434CrossRefPubMedGoogle Scholar
  26. Rome LH, Kedersha N, Chugani D (1991) Unlocking vaults: organelles in search of a function. Trends Cell Biol 1:47–50CrossRefPubMedGoogle Scholar
  27. Scheffer GL, Wijngaard PLJ, Flens MJ, Izquierdo MA, Slovak ML, Pinedo HM, Meijer CJLM, Clevers HC, Scheper RJ (1995) The drug resistance-related protein LRP is the human major vault protein. Nature Med 1:578–582CrossRefPubMedGoogle Scholar
  28. Schroeijers AB, Siva AC, Scheffer GL, Jong MC de, Bolick SCE, Dukers DF, Slootstra JW, Meloen RH, Wiemer E, Kickhoefer VA, Rome LH, Scheper RJ (2000) The Mr 193,000 vault protein is up-regulated in multidrug-resistant cancer cell lines. Cancer Res 60:1104–1110PubMedGoogle Scholar
  29. Suprenant KA (2002) Vault ribonucleoprotein particles: sarcophagi, gondolas, or safety deposit boxes. Biochemistry 41:14447–14454CrossRefPubMedGoogle Scholar
  30. Stephen AG, Raval-Fernandez S, Huyn T, Torres M, Kickhoefer VA, Rome LH (2001) Assembly of vault-like particles in insect cells expressing only the major vault protein. J Biol Chem 276:23217–23220CrossRefPubMedGoogle Scholar
  31. Trotman LC, Mosberger N, Fornerod M, Stidwill RP, Greber UF (2001) Import of adenovirus DNA involves the nuclear pore complex receptor CAN/Nup214 and histone H1. Nature Cell Biol 3:1092–1100CrossRefPubMedGoogle Scholar
  32. Van Zon A, Mossine MH, Scheper RJ, Sonneveld P, Wiemer EAC (2003) The vault complex. Cell Mol Life Sci 60:1828–1837CrossRefPubMedGoogle Scholar
  33. Zhang L, Volknandt W, Gundelfinger ED, Zimmermann H (2000) A comparison of synaptic protein localization in hippocampal mossy fiber nerve terminals and neurosecretory endings of the neurohypophysis using the cryo-immunogold technique. J Neurocytol 29:19–30CrossRefPubMedGoogle Scholar

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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