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© The Rockefeller University Press, 0021-9525/1999//1163 $5.00
The Journal of Cell Biology, Volume 144, Number 6, , 1999 1163-1172

Recombinant Major Vault Protein Is Targeted to Neuritic Tips of PC12 Cells

Christine Herrmann^*, Elaheh Golkaramnay^*, Elisabeth Inman, Leonard Rome, and Walter Volknandt^*

^* Biozentrum der J. W. Goethe-Universität, D-60439 Frankfurt/Main, Germany; and Department of Biological Chemistry, University of California, Los Angeles School of Medicine, Los Angeles, California 90024-1737

The major vault protein (MVP) is the predominant constituent of ubiquitous, evolutionarily conserved large cytoplasmic ribonucleoprotein particles of unknown function. Vaults are multimeric protein complexes with several copies of an untranslated RNA. Double labeling employing laser-assisted confocal microscopy and indirect immunofluorescence demonstrates partial colocalization of vaults with cytoskeletal elements in Chinese hamster ovary (CHO) and nerve growth factor (NGF)-treated neuronlike PC12 cells. Transfection of CHO and PC12 cells with a cDNA encoding the rat major vault protein containing a vesicular stomatitis virus glycoprotein epitope tag demonstrates that the recombinant protein is sorted into vault particles and targeted like endogenous MVPs. In neuritic extensions of differentiated PC12 cells, there is an almost complete overlap of the distribution of microtubules and vaults. A pronounced colocalization of vaults with filamentous actin can be seen in the tips of neurites. Moreover, in NGF-treated PC12 cells the location of vaults partially coincides with vesicular markers. Within the terminal tips of neurites vaults are located near secretory organelles. Our observations suggest that the vault particles are transported along cytoskeletal-based cellular tracks.

Key Words: vaults • protein targeting • major vault protein • transport • PC12

Abbreviations used in this paper: GLUT4, glucose transporter 4; LRP, lung resistance–related protein; MVP, major vault protein; pvMVP, plasmid encoding VSVG-tagged MVP; SV2, synaptical vesicle protein 2; vMVP, VSVG-tagged MVP; vRNA, vault RNA; VSVG, vesicular stomatitis virus glycoprotein.

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