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Updated: Sep 26, 2026

A Cell Free Assay to Study Chromatin Decondensation at the End of Mitosis
Published on: December 19, 2015
Removal of a single pore subcomplex results in vertebrate nuclei devoid of nuclear pores
Amnon Harel1, Arturo V Orjalo, Thomas Vincent
1Section of Cell and Developmental Biology, Division of Biology 0347, University of California, San Diego, La Jolla 92093, USA.
Abstract:
The vertebrate nuclear pore complex, 30 times the size of a ribosome, assembles from a library of soluble subunits and two membrane proteins. Using immunodepletion of Xenopus nuclear reconstitution extracts, it has previously been possible to assemble nuclei lacking pore subunits tied to protein import, export, or mRNA export. However, these altered pores all still possessed the bulk of pore structure. Here, we immunodeplete a single subunit, the Nup107-160 complex, using antibodies to Nup85 and Nup133, two of its components. The resulting reconstituted nuclei are severely defective for NLS import and DNA replication. Strikingly, they show a profound defect for every tested nucleoporin. Even the integral membrane proteins POM121 and gp210 are absent or unorganized. Scanning electron microscopy reveals pore-free nuclei, while addback of the Nup107-160 complex restores functional pores. We conclude that the Nup107-160 complex is a pivotal determinant for vertebrate nuclear pore complex assembly.
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