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PMID:11387327

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Citation

Allen, NP, Huang, L, Burlingame, A and Rexach, M (2001) Proteomic analysis of nucleoporin interacting proteins. J. Biol. Chem. 276:29268-74

Abstract

The Saccharomyces cerevisiae nuclear pore complex is a supramolecular assembly of 30 nucleoporins that cooperatively facilitate nucleocytoplasmic transport. Thirteen nucleoporins that contain FG peptide repeats (FG Nups) are proposed to function as stepping stones in karyopherin-mediated transport pathways. Here, protein interactions that occur at individual FG Nups were sampled using immobilized nucleoporins and yeast extracts. We find that many proteins bind to FG Nups in highly reproducible patterns. Among 135 proteins identified by mass spectrometry, most were karyopherins and nucleoporins. The PSFG nucleoporin Nup42p and the GLFG nucleoporins Nup49p, Nup57p, Nup100p, and Nup116p exhibited generic interactions with karyopherins; each bound 6--10 different karyopherin betas, including importins as well as exportins. Unexpectedly, the same Nups also captured the hexameric Nup84p complex and Nup2p. In contrast, the FXFG nucleoporins Nup1p, Nup2p, and Nup60p were more selective and captured mostly the Kap95p.Kap60p heterodimer. When the concentration of Gsp1p-GTP was elevated in the extracts to mimic the nucleoplasmic environment, the patterns of interacting proteins changed; exportins exhibited enhanced binding to FG Nups, and importins exhibited reduced binding. The results demonstrate a global role for Gsp1p-GTP on karyopherin-nucleoporin interactions and provide a rudimentary map of the routes that karyopherins take as they cross the nuclear pore complex.

Links

PubMed Online version:10.1074/jbc.M102629200

Keywords

Cell Nucleus/metabolism; Cytoplasm/metabolism; Dimerization; Fungal Proteins/chemistry; Fungal Proteins/isolation & purification; Fungal Proteins/metabolism; Macromolecular Substances; Membrane Proteins/chemistry; Membrane Proteins/isolation & purification; Membrane Proteins/metabolism; Nuclear Proteins/chemistry; Nuclear Proteins/isolation & purification; Nuclear Proteins/metabolism; Protein Transport; Proteome/chemistry; Proteome/isolation & purification; Proteome/metabolism; Recombinant Proteins/chemistry; Recombinant Proteins/metabolism; Repetitive Sequences, Amino Acid; Saccharomyces cerevisiae/metabolism; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

YEAST:NUP2

enables

GO:0017056: structural constituent of nuclear pore

ECO:0000316: genetic interaction evidence used in manual assertion

SGD:S000004284
SGD:S000004339

F

Seeded From UniProt

complete

YEAST:NUP57

enables

GO:0017056: structural constituent of nuclear pore

ECO:0000316: genetic interaction evidence used in manual assertion

SGD:S000000912
SGD:S000002743
SGD:S000002803
SGD:S000003450
SGD:S000004284
SGD:S000004339
SGD:S000004925
SGD:S000006046

F

Seeded From UniProt

complete

YEAST:NUP42

enables

GO:0017056: structural constituent of nuclear pore

ECO:0000316: genetic interaction evidence used in manual assertion

SGD:S000000912
SGD:S000002743
SGD:S000003450
SGD:S000004284
SGD:S000004339
SGD:S000004848
SGD:S000006046

F

Seeded From UniProt

complete

YEAST:NUP49

enables

GO:0017056: structural constituent of nuclear pore

ECO:0000316: genetic interaction evidence used in manual assertion

SGD:S000000912
SGD:S000002743
SGD:S000003450
SGD:S000004284
SGD:S000004339
SGD:S000004925
SGD:S000006046

F

Seeded From UniProt

complete

YEAST:NU116

enables

GO:0017056: structural constituent of nuclear pore

ECO:0000316: genetic interaction evidence used in manual assertion

SGD:S000000221
SGD:S000000912
SGD:S000002743
SGD:S000002803
SGD:S000002984
SGD:S000003450
SGD:S000004284
SGD:S000004339
SGD:S000004925
SGD:S000005686
SGD:S000006046

F

Seeded From UniProt

complete

YEAST:NUP60

enables

GO:0017056: structural constituent of nuclear pore

ECO:0000316: genetic interaction evidence used in manual assertion

SGD:S000000912
SGD:S000004284
SGD:S000004339

F

Seeded From UniProt

complete


See also

References

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