The entire Nup107-160 complex, including three new members, is targeted as one entity to kinetochores in mitosis.
In eukaryotes, bidirectional transport of macromolecules between the cytoplasm and the nucleus occurs through elaborate supramolecular structures embedded in the nuclear envelope, the nuclear pore complexes (NPCs). NPCs are composed of multiple copies of approximately 30 different proteins termed nucleoporins, of which several can be biochemically isolated as subcomplexes. One such building block of the NPC, termed the Nup107-160 complex in vertebrates, was so far demonstrated to be composed of six different nucleoporins. Here, we identify three WD (Trp-Asp)-repeat nucleoporins as new members of this complex, two of which, Nup37 and Nup43, are specific to higher eukaryotes. The third new member Seh1 is more loosely associated with the Nup107-160 complex biochemically, but its depletion by RNA interference leads to phenotypes similar to knock down of other constituents of this complex. By combining green fluorescent protein-tagged nucleoporins and specific antibodies, we show that all the constituents of this complex, including Nup37, Nup43, Seh1, and Sec13, are targeted to kinetochores from prophase to anaphase of mitosis. Together, our results indicate that the entire Nup107-160 complex, which comprises nearly one-third of the so-far identified nucleoporins, specifically localizes to kinetochores in mitosis.
Loïodice
Isabelle
I
Unité Mixte de Recherche 144 Centre National de la Recherche Scientifique-Institut Curie, Section Recherche, 75248 Paris Cedex 05, France.
Alves
Annabelle
A
Rabut
Gwénaël
G
Van Overbeek
Megan
M
Ellenberg
Jan
J
Sibarita
Jean-Baptiste
JB
Doye
Valérie
V
D000818:Animals; D022003:Cell Nucleus Structures / Q000378:metabolism; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D018386:Kinetochores / Q000378:metabolism*; D008938:Mitosis; D028861:Nuclear Pore Complex Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference
C000605772:NUP43 protein, human; D028861:Nuclear Pore Complex Proteins