Regular Article
Structural Domains of Vault Proteins: A Role for the Coiled Coil Domain in Vault Assembly☆

https://doi.org/10.1006/bbrc.2002.6472Get rights and content

Abstract

Vaults consist of multiple copies of three proteins (MVP, VPARP, and TEP1) and several untranslated RNAs. The function of vaults is unknown but the typical and evolutionary conserved structure indicates a role in intracellular transport. Although all vault components have been identified and characterized, not much is known about vault protein assembly. In this study we identified and analyzed structural domains involved in vault assembly with emphasis on protein–protein interactions. Using a yeast two-hybrid system, we demonstrate within MVP an intramolecular binding site and show that MVP molecules interact with each other via their coiled coil domain. We show that purified MVP is able to bind calcium, most likely at calcium-binding EF-hands. No interactions could be detected between TEP1 and other vault proteins. However, the N-terminal half of MVP binds to a specific domain in the C-terminus of VPARP. Furthermore, VPARP contains amino acid stretches mediating intramolecular binding.

References (33)

  • U.K. Laemmli et al.

    Maturation of the head of bacteriophage T4. I. DNA packaging events

    J. Mol. Biol.

    (1973)
  • A.G. Stephen et al.

    Assembly of vault-like particles in insect cells expressing only the major vault protein

    J. Biol. Chem.

    (2001)
  • C. Abbondanza et al.

    Interaction of vault particles with estrogen receptor in the MCF-7 breast cancer cell

    J. Cell. Biol.

    (1998)
  • D.C. Chugani et al.

    Evidence that vault ribonucleoprotein particles localize to the nuclear pore complex

    J. Cell Sci.

    (1993)
  • N.L. Kedersha et al.

    Isolation and characterization of a novel ribonucleoprotein particle: Large structures contain a single species of small RNA

    J. Cell. Biol.

    (1986)
  • C. Herrmann et al.

    Recombinant major vault protein is targeted to neuritic tips of PC12 cells

    J. Cell Biol.

    (1999)

    • Cited by (0)

    ☆

    Abbreviations used: AD, transactivation domain; BD, DNA-binding domain; BRCT, BRCA1 C-terminus; MVP, major vault protein; TEP1, telomerase associated protein; VPARP, vault poly(ADP-ribose) polymerase.

    1

    To whom correspondence and reprint requests should be addressed at Institute of Hematology, Erasmus University, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands. Fax: (+31)-10-408 9470. E-mail: [email protected].

    View full text
    Copyright © 2002 Elsevier Science (USA). All rights reserved.