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

  1. Author:
    Stephen, A. G.
    Raval-Fernandes, S.
    Huynh, T.
    Torres, M.
    Kickhoefer, V. A.
    Rome, L. H.

  2. Author Address

    Univ Calif Los Angeles, Sch Med, Dept Biol Chem, 33-131 CHS, 10833 Le Conte Ave, Los Angeles, CA 90095 USA. Univ Calif Los Angeles, Sch Med, Dept Biol Chem, Los Angeles, CA 90095 USA. Univ Calif Los Angeles, Sch Med, Jonsson Comprehens Canc Ctr, Los Angeles, CA 90095 USA. NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, Frederick, MD 21702 USA. Rome LH Univ Calif Los Angeles, Sch Med, Dept Biol Chem, 33-131 CHS, 10833 Le Conte Ave, Los Angeles, CA 90095 USA.
    1. Year: 2001
  2. Journal: Journal of Biological Chemistry
    1. 276
    2. 26
    3. Pages: 23217-23220
  4. Type of Article: Article
  1. Abstract:

    Vaults are the largest (13 megadalton) cytoplasmic ribonucleoprotein particles known to exist in eukaryotic cells. They have a unique barrel-shaped structure with 8-fold symmetry. Although the precise function of vaults is unknown, their wide distribution and highly conserved morphology in eukaryotes suggests that their function is essential and that their structure must be important for their function. The 100- kDa major vault protein (MVP) constitutes similar to 75% of the particle mass and is predicted to form the central barrel portion of the vault. To gain insight into the mechanisms for vault assembly, we have expressed rat MVP in the Sf9 insect cell line using a baculovirus vector. Our results show that the expression of the rat MVP alone can direct the formation of particles that have biochemical characteristics similar to endogenous rat vaults and display the distinct vault-like morphology when negatively stained and examined by electron microscopy. These particles are the first example of a single protein polymerizing into a non-spherically, non-cylindrically symmetrical structure. Understanding vault assembly will enable us to design agents that disrupt vault formation and hence aid in elucidating vault function in vivo.

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