The formation of vault-tubes: a dynamic interaction between vaults and vault PARP
Vaults are barrel-shaped cytoplasmic ribonucleoprotein particles that are composed of a major vault protein (MVP), two minor vault proteins [telomerase-associated protein 1 (TEP1), vault poly(ADP-ribose) polymerase (VPARP)] and small untranslated RNA molecules. Not all expressed TEP1 and VPARP in cells is bound to vaults. TEP1 is known to associate with the telomerase complex, whereas VPARP is also present in the nuclear matrix and in cytoplasmic clusters (VPARP-rods). We examined the subcellular localization and the dynamics of the vault complex in a non-small cell lung cancer cell line expressing MVP tagged with green fluorescent protein. Using quantitative fluorescence recovery after photobleaching (FRAP) it was shown that vaults move temperature independently by diffusion. However, incubation at room temperature (21 degrees C) resulted in the formation of distinct tube-like structures in the cytoplasm. Raising the temperature could reverse this process. When the vault-tubes were formed, there were fewer or no VPARP-rods present in the cytoplasm, suggesting an incorporation of the VPARP into the vault-tubes. MVP molecules have to interact with each other via their coiled-coil domain in order to form vault-tubes. Furthermore, the stability of microtubules influenced the efficiency of vault-tube formation at 21 degrees C. The dynamics and structure of the tubes were examined using confocal microscopy. Our data indicate a direct and dynamic relationship between vaults and VPARP, providing further clues to unravel the function of vaults.
|Keywords||Fluorescence Recovery After Photobleaching, Hela Cells, Humans, Microtubules/*metabolism, Poly(ADP-ribose) Polymerases/*metabolism, Protein Binding, Protein Structure, Tertiary, Protein Transport, Recombinant Fusion Proteins/metabolism, Research Support, Non-U.S. Gov't, Subcellular Fractions, Tumor Cells, Cultured, Vault Ribonucleoprotein Particles/*metabolism|
van Zon, A., Mossink, M.H., Schoester, M., Houtsmuller, A.B., Scheffer, G.L., Scheper, R.J., … Wiemer, E.A.C.. (2003). The formation of vault-tubes: a dynamic interaction between vaults and vault PARP. Journal of Cell Science. Retrieved from http://hdl.handle.net/1765/8363