Nup50, a nucleoplasmically oriented nucleoporin with a role in nuclear protein export.
We present here a detailed analysis of a rat polypeptide termed Nup50 (formerly NPAP60) that was previously found to be associated with the nuclear pore complex (F. Fan et al., Genomics 40:444-453, 1997). We have found that Nup50 (and/or a related 70-kDa polypeptide) is present in numerous rat cells and tissues. By immunofluorescence microscopy, Nup50 was found to be highly concentrated at the nuclear envelope of rat liver nuclei, whereas in cultured NRK cells it also is abundant in intranuclear regions. On the basis of immunogold electron microscopy of both rat liver nuclear envelopes and NRK cells, we determined that Nup50 is specifically localized in the nucleoplasmic fibrils of the pore complex. Microinjection of anti-Nup50 antibodies into the nucleus of NRK cells resulted in strong inhibition of nuclear export of a protein containing a leucine-rich nuclear export sequence, whereas nuclear import of a protein containing a classical nuclear localization sequence was unaffected. Correspondingly, CRM1, the export receptor for leucine-rich export sequences, directly bound to a fragment of Nup50 in vitro, whereas several other import and export receptors did not significantly interact with this fragment. Taken together, our data indicate that Nup50 has a direct role in nuclear protein export and probably serves as a binding site on the nuclear side of the pore complex for export receptor-cargo complexes.
Guan
T
T
Departments of Cell and Molecular Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
Kehlenbach
R H
RH
Schirmer
E C
EC
Kehlenbach
A
A
Fan
F
F
Clurman
B E
BE
Arnheim
N
N
Gerace
L
L
D000818:Animals; D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D029563:Cellular Apoptosis Susceptibility Protein; D028884:Karyopherins; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008856:Microscopy, Fluorescence; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins / Q000378:metabolism*; D018272:Porins / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear; D000097606:Exportin 1 Protein
D002352:Carrier Proteins; D029563:Cellular Apoptosis Susceptibility Protein; D028884:Karyopherins; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D018272:Porins; D011506:Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D000097606:Exportin 1 Protein; C484544:Nup50 protein, rat