Assembly of nuclear pore complexes mediated by major vault protein.
During interphase growth of eukaryotic cells, nuclear pore complexes (NPCs) are continuously incorporated into the intact nuclear envelope (NE) by mechanisms that are largely unknown. De novo formation of NPCs involves local fusion events between the inner and outer nuclear membrane, formation of a transcisternal membranous channel of defined diameter and the coordinated assembly of hundreds of nucleoporins into the characteristic NPC structure. Here we have used a cell-free system based on Xenopus egg extract, which allows the experimental separation of nuclear-membrane assembly and NPC formation. Nuclei surrounded by a closed double nuclear membrane, but devoid of NPCs, were first reconstituted from chromatin and a specific membrane fraction. Insertion of NPCs into the preformed pore-free nuclei required cytosol containing soluble nucleoporins or nucleoporin subcomplexes and, quite unexpectedly, major vault protein (MVP). MVP is the main component of vaults, which are ubiquitous barrel-shaped particles of enigmatic function. Our results implicate MVP, and thus also vaults, in NPC biogenesis and provide a functional explanation for the association of a fraction of vaults with the NE and specifically with NPCs in intact cells.
Vollmar
Friederike
F
Division of Electron Microscopy, Biocenter, University of Würzburg, 97074 Würzburg, Germany.
Hacker
Christian
C
Zahedi
René-Peiman
RP
Sickmann
Albert
A
Ewald
Andrea
A
Scheer
Ulrich
U
Dabauvalle
Marie-Christine
MC
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D015151:Immunoblotting; D022022:Nuclear Pore / Q000378:metabolism* / Q000648:ultrastructure; D010063:Ovum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D020394:Vault Ribonucleoprotein Particles / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure; D014982:Xenopus laevis / Q000378:metabolism*; D000099187:Major Vault Protein
D000906:Antibodies; D011994:Recombinant Proteins; D020394:Vault Ribonucleoprotein Particles; D000099187:Major Vault Protein