The conserved Nup107-160 complex is critical for nuclear pore complex assembly.
Nuclear pore complexes (NPCs) are large multiprotein assemblies that allow traffic between the cytoplasm and the nucleus. During mitosis in higher eukaryotes, the Nuclear Envelope (NE) breaks down and NPCs disassemble. How NPCs reassemble and incorporate into the NE upon mitotic exit is poorly understood. We demonstrate a function for the conserved Nup107-160 complex in this process. Partial in vivo depletion of Nup133 or Nup107 via RNAi in HeLa cells resulted in reduced levels of multiple nucleoporins and decreased NPC density in the NE. Immunodepletion of the entire Nup107-160 complex from in vitro nuclear assembly reactions produced nuclei with a continuous NE but no NPCs. This phenotype was reversible only if Nup107-160 complex was readded before closed NE formation. Depletion also prevented association of FG-repeat nucleoporins with chromatin. We propose a stepwise model in which postmitotic NPC assembly initiates on chromatin via early recruitment of the Nup107-160 complex.
Walther
Tobias C
TC
EMBL, Meyerhofstrasse 1, 69117 Heidelberg, Germany.
Alves
Annabelle
A
Pickersgill
Helen
H
Loïodice
Isabelle
I
Hetzer
Martin
M
Galy
Vincent
V
Hülsmann
Bastian B
BB
Köcher
Thomas
T
Wilm
Matthias
M
Allen
Terry
T
Mattaj
Iain W
IW
Doye
Valérie
V
D000818:Animals; D002457:Cell Extracts; D002614:Chelating Agents / Q000494:pharmacology; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D005057:Eukaryotic Cells / Q000378:metabolism* / Q000648:ultrastructure; D005260:Female; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D015778:Minor Histocompatibility Antigens; D009685:Nuclear Envelope / Q000378:metabolism* / Q000648:ultrastructure; D022022:Nuclear Pore / Q000378:metabolism* / Q000648:ultrastructure; D028861:Nuclear Pore Complex Proteins / Q000172:deficiency* / Q000235:genetics; D009687:Nuclear Proteins; D009865:Oocytes; D010641:Phenotype; D029867:Xenopus Proteins; D014982:Xenopus laevis
D002457:Cell Extracts; D002614:Chelating Agents; D002843:Chromatin; D015778:Minor Histocompatibility Antigens; C512772:NUP107 protein, human; C440250:NUP133 protein, human; C440251:NUP160 protein, human; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; C499080:Nup160 protein, Xenopus; D029867:Xenopus Proteins