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dc.contributor.authorHo, Alberto K
dc.contributor.authorShen, Tian X
dc.contributor.authorRyan, Kathryn J
dc.contributor.authorKiseleva, Elena
dc.contributor.authorLevy, Marilyn A
dc.contributor.authorAllen, Terence D
dc.contributor.authorWente, Susan R
dc.date.accessioned2009-11-18T10:33:33Z
dc.date.available2009-11-18T10:33:33Z
dc.date.issued2000-08
dc.identifier.citationAssembly and preferential localization of Nup116p on the cytoplasmic face of the nuclear pore complex by interaction with Nup82p. 2000, 20 (15):5736-48 Mol. Cell. Biol.en
dc.identifier.issn0270-7306
dc.identifier.pmid10891509
dc.identifier.urihttp://hdl.handle.net/10541/86376
dc.description.abstractThe yeast Saccharomyces cerevisiae nucleoporin Nup116p serves as a docking site for both nuclear import and export factors. However, the mechanism for assembling Nup116p into the nuclear pore complex (NPC) has not been resolved. By conducting a two-hybrid screen with the carboxy (C)-terminal Nup116p region as bait, we identified Nup82p. The predicted coiled-coil region of Nup82p was not required for Nup116p interaction, making the binding requirements distinct from those for the Nsp1p-Nup82p-Nup159p subcomplex (N. Belgareh, C. Snay-Hodge, F. Pasteau, S. Dagher, C. N. Cole, and V. Doye, Mol. Biol. Cell 9:3475-3492, 1998). Immunoprecipitation experiments using yeast cell lysates resulted in the coisolation of a Nup116p-Nup82p subcomplex. Although the absence of Nup116p had no effect on the NPC localization of Nup82p, overexpression of C-terminal Nup116p in a nup116 null mutant resulted in Nup82p mislocalization. Moreover, NPC localization of Nup116p was specifically diminished in a nup82-Delta108 mutant after growth at 37 degrees C. Immunoelectron microscopy analysis showed Nup116p was localized on both the cytoplasmic and nuclear NPC faces. Its distribution was asymmetric with the majority at the cytoplasmic face. Taken together, these results suggest that Nup82p and Nup116p interact at the cytoplasmic NPC face, with nucleoplasmic Nup116p localization utilizing novel binding partners.
dc.language.isoenen
dc.subject.meshCell Nucleus
dc.subject.meshCytoplasm
dc.subject.meshFluorescent Antibody Technique
dc.subject.meshFungal Proteins
dc.subject.meshGreen Fluorescent Proteins
dc.subject.meshLuminescent Proteins
dc.subject.meshMembrane Proteins
dc.subject.meshMutation
dc.subject.meshNuclear Pore Complex Proteins
dc.subject.meshNuclear Proteins
dc.subject.meshRecombinant Proteins
dc.subject.meshSaccharomyces cerevisiae
dc.subject.meshSaccharomyces cerevisiae Proteins
dc.subject.meshTemperature
dc.subject.meshTwo-Hybrid System Techniques
dc.titleAssembly and preferential localization of Nup116p on the cytoplasmic face of the nuclear pore complex by interaction with Nup82p.en
dc.typeArticleen
dc.contributor.departmentDepartment of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.en
dc.identifier.journalMolecular and Cellular Biologyen
html.description.abstractThe yeast Saccharomyces cerevisiae nucleoporin Nup116p serves as a docking site for both nuclear import and export factors. However, the mechanism for assembling Nup116p into the nuclear pore complex (NPC) has not been resolved. By conducting a two-hybrid screen with the carboxy (C)-terminal Nup116p region as bait, we identified Nup82p. The predicted coiled-coil region of Nup82p was not required for Nup116p interaction, making the binding requirements distinct from those for the Nsp1p-Nup82p-Nup159p subcomplex (N. Belgareh, C. Snay-Hodge, F. Pasteau, S. Dagher, C. N. Cole, and V. Doye, Mol. Biol. Cell 9:3475-3492, 1998). Immunoprecipitation experiments using yeast cell lysates resulted in the coisolation of a Nup116p-Nup82p subcomplex. Although the absence of Nup116p had no effect on the NPC localization of Nup82p, overexpression of C-terminal Nup116p in a nup116 null mutant resulted in Nup82p mislocalization. Moreover, NPC localization of Nup116p was specifically diminished in a nup82-Delta108 mutant after growth at 37 degrees C. Immunoelectron microscopy analysis showed Nup116p was localized on both the cytoplasmic and nuclear NPC faces. Its distribution was asymmetric with the majority at the cytoplasmic face. Taken together, these results suggest that Nup82p and Nup116p interact at the cytoplasmic NPC face, with nucleoplasmic Nup116p localization utilizing novel binding partners.


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