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PMID:9813081

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Citation

Fahrenkrog, B, Hurt, EC, Aebi, U and Panté, N (1998) Molecular architecture of the yeast nuclear pore complex: localization of Nsp1p subcomplexes. J. Cell Biol. 143:577-88

Abstract

The nuclear pore complex (NPC), a supramolecular assembly of approximately 100 different proteins (nucleoporins), mediates bidirectional transport of molecules between the cytoplasm and the cell nucleus. Extensive structural studies have revealed the three- dimensional (3D) architecture of Xenopus NPCs, and eight of the approximately 12 cloned and characterized vertebrate nucleoporins have been localized within the NPC. Thanks to the power of yeast genetics, 30 yeast nucleoporins have recently been cloned and characterized at the molecular level. However, the localization of these nucleoporins within the 3D structure of the NPC has remain elusive, mainly due to limitations of preparing yeast cells for electron microscopy (EM). We have developed a new protocol for preparing yeast cells for EM that yielded structurally well-preserved yeast NPCs. A direct comparison of yeast and Xenopus NPCs revealed that the NPC structure is evolutionarily conserved, although yeast NPCs are 15% smaller in their linear dimensions. With this preparation protocol and yeast strains expressing nucleoporins tagged with protein A, we have localized Nsp1p and its interacting partners Nup49p, Nup57p, Nup82p, and Nic96p by immuno-EM. Accordingly, Nsp1p resides in three distinct subcomplexes which are located at the entry and exit of the central gated channel and at the terminal ring of the nuclear basket.

Links

PubMed PMC2148146

Keywords

Animals; Calcium-Binding Proteins; Cell Nucleus/chemistry; Cytoplasm/chemistry; Fungal Proteins/analysis; Green Fluorescent Proteins; Luminescent Proteins/analysis; Membrane Proteins/analysis; Nuclear Envelope/chemistry; Nuclear Pore Complex Proteins; Nuclear Proteins/analysis; Recombinant Fusion Proteins/analysis; Saccharomyces cerevisiae/chemistry; Saccharomyces cerevisiae/ultrastructure; Saccharomyces cerevisiae Proteins; Xenopus

Significance

Annotations

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

YEAST:NSP1

part_of

GO:0044613: nuclear pore central transport channel

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

YEAST:NSP1

part_of

GO:0044615: nuclear pore nuclear basket

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

YEAST:NIC96

part_of

GO:0044615: nuclear pore nuclear basket

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

YEAST:NUP57

part_of

GO:0044613: nuclear pore central transport channel

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

YEAST:NUP49

part_of

GO:0044613: nuclear pore central transport channel

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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